Cryptococci

Clin Immunol, 1999 Feb, 90(2), 256 - 65
Involvement of nitric oxide in protecting mechanism during experimental cryptococcosis; Rossi GR et al.; In the present study we investigated the role of nitric oxide (NO) in the effector mechanisms of host defense against Cryptococcus neoformans in vivo . Our results showed an increase of NO produced by the peritoneal macrophages from 14-days infected rats compared with normal rats . These cells were capable of killing C . neoformans to a greater extent than macrophages from noninfected rats (80% vs 20%, respectively) . The killing of C . neoformans by infected cells was efficiently inhibited (80% to 35%, P < 0.001) by adding aminoguanidine (AG) to the cultures . We observed that in vivo administration of AG to the infected animals efficiently inhibited the metabolism producing NO and failed to affect that of normal animals . When the NO synthase (NOS) was inhibited in vivo in the infected animals, a marked increase of the fungi charge in the organs was observed with respect to the normal animals treated with AG . We also observed that the course of the infection is drastically modified after the inhibition of NO production because all the animals infected and treated with AG died from cryptococcosis before 20 days postinfection (p.i.) . These results indicate that NO is a crucial molecule in the effector mechanisms in this infection model .

J Clin Oncol, 1999 Feb, 17(2), 554 - 60
Value of combined approach with thallium-201 single-photon emission computed tomography and Epstein-Barr virus DNA polymerase chain reaction in CSF for the diagnosis of AIDS-related primary CNS lymphoma; Antinori A et al.; PURPOSE: To determine the diagnostic capability of thallium-201 (201Tl) single-photon emission computed tomography (SPECT) combined with Epstein-Barr virus DNA (EBV-DNA) in CSF for the diagnosis of AIDS-related primary CNS lymphoma (PCNSL) . PATIENTS AND METHODS: All human immunodeficiency virus (HIV)-infected patients with focal brain lesions observed between June 1996 and March 1998 underwent lumbar puncture and 201Tl SPECT . Each CSF sample was tested with polymerase chain reaction (PCR) for EBV-DNA . RESULTS: Thirty-one patients were included, 13 with PCNSL and 18 with nontumor disorders . In 11 PCNSL patients, EBV-DNA was positive . Thallium-201 uptake ranged from 1.90 to 4.07 in PCNSL cases (mean, 2.77; 95% confidence interval {CI}, 2.35 to 3.19) and from 0.91 to 3.38 in nontumor patients (mean, 1.62; 95% CI, 1.30 to 1.94) (P<.0002) . Using a lesion/background ratio of 1.95 as cutoff, a negative SPECT was found in one PCNSL case and 16 nonneoplastic cases . A cryptococcoma and a tuberculoma showed highly increased 201Tl uptake . Epstein-Barr virus DNA was never detected in nonneoplastic patients . For PCNSL diagnosis, hyperactive lesions showed 92% sensitivity and 94% negative predictive value (NPV), whereas positive EBV-DNA had 100% specificity and 100% positive predictive value . The presence of increased uptake and/or positive EBV-DNA had 100% sensitivity and 100% NPV . CONCLUSION: Combined SPECT and EBV-DNA showed a very high diagnostic accuracy for AIDS-related PCNSL . Because PCNSL likelihood is extremely high in patients with hyperactive lesions and positive EBV-DNA, brain biopsy could be avoided, and patients could promptly undergo radiotherapy or multimodal therapy . On the contrary, in patients showing hypoactive lesions with negative EBV-DNA, empiric anti-Toxoplasma therapy is indicated . In patients with discordant SPECT/PCR results, brain biopsy seems to be advisable.

Rinsho Shinkeigaku, 1998 Sep, 38(9), 831 - 7
{Serial MRI findings in patients with CNS cryptococcosis}; Kumazawa K et al.; We reported the serial magnetic resonance imaging (MRI) findings of two patients with central nervous system (CNS) cryptococcal infection without AIDS . The diagnosis of CNS cryptococcosis was made by visualizing the fungi in the CSF with the India ink test, detecting cryptococcal antigens, and culturing the fungus . Both patients had dilated perivascular Virchow-Robin (V-R) spaces, which were defined as small rounded lesions greater less than 3mm diameter that were hyperintense on T2-weighted images . They were present in the basal ganglia, brainstem and cerebral white matter . Case 1 had bilateral parietal arachnoid cyst which was thought to represent a focal collection of organisms and mucoid material within subarachnoid space . Abnormal optochiasmatic arachnoid enhancement detected in case 2, who had complete loss of vision . With disease progression perivascular V-R increased in size, resulting in the developing cryptococomas which were defined as rounded lesions greater than 3mm diameter, and were hyperintense on T2-weighted images in the basal ganglia, cerebellum and cerebral white matter . In follow-up MRI of those patients, radiological progression was seen despite appropriate treatment and falling CSF cryptococcal antigens . In conclusion, this spectrum of MRI appearances in CNS cryptococcosis reflects the pathological mechanism of invasion by the fungus, and may be relatively specific for cryptococcosis.

Appl Microbiol Biotechnol, 1999 Jan, 51(1), 40 - 5
Production of sophorolipids from whey: development of a two-stage process with Cryptococcus curvatus ATCC 20509 and Candida bombicola ATCC 22214 using deproteinized whey concentrates as substrates; Daniel HJ et al.; In order to produce sophorolipids from whey, thereby lowering the lactose content and biological oxygen demand, a two-step batch cultivation process was developed including medium sterilization by filtration . In the first step, whey was sterilized by a combination of crossflow and sterile filtration . Because the sophorolipid-producing yeast Candida bombicola ATCC 22214 was not able to use lactose as a carbon source directly, the oleaginous yeast Cryptococcus curvatus ATCC 20509 was grown on deproteinized whey concentrates (DWC) . With 1:1 diluted DWC-20, lactose was consumed as the carbon source and biomass (24 g/l dry weight content) as well as single-cell oil (SCO, 10 g/l) were produced . The cultivation broth was disrupted with a glass bead mill and it served as medium for growth (29 g cell dry mass/l) and sophorolipid production (12 g/l) of the yeast C . bombicola.

Med Mycol, 1998 Oct, 36(5), 341 - 4
First isolation of Cryptococcus neoformans var . gattii, serotype C, from the environment in Colombia; Callejas A et al.; The natural habitat of Cryptococcus neoformans var . gattii, serotype B in the environment was established by Australian investigators who demonstrated its association with species of Eucalyptus . The aim of the present study was to search for the habitat of this variety in a city of Colombia, where clinical cases due to this variety occur with great frequency . For a period of 5 months detritus, vegetable material and air samples in and around 68 almond trees (Terminalia catappa) located in the city were studied . C . neoformans var . gattii serotype C was the only variety isolated from two of the 68 trees sampled . These trees were positive for 4 of the 5 months during which they were studied . From the first positive sample kept under refrigeration, it was possible to isolate the fungus up to 3 months later . This is the first report of the isolation of serotype C from the environment . More studies are required in order to establish the ecological significance of this finding.

Med Mycol, 1998 Oct, 36(5), 335 - 9
First report of Cryptococcus laurentii meningitis and a fatal case of Cryptococcus albidus cryptococcaemia in AIDS patients; Kordossis T et al.; We report the first case of Cryptococcus laurentii meningitis and a rare case of Cryptococcus albidus cryptococcaemia in AIDS patients . Both infections were treated with amphotericin B and flucytosine . The C . laurentii meningitis was controlled after 2 weeks of treatment with no evidence of infection 20 months later . The patient with C . albidus cryptococcaemia, despite the amphotericin B/flucytosine combination therapy, died on the 14th day of treatment . The minimum inhibitory concentrations (MICs) for C . laurentii, as determined by Etest on RPMI 1640 agar, were 0.25 microg ml(-1) of amphotericin B, 1.25 microg ml(-1) flucytosine, 4 microg ml(-1) fluconazole, 0.50 microg ml(-1) itraconazole and 1.0 microg ml(-1) of ketoconazole . The MIC of amphotericin B for C . albidus was 0.5 microg ml(-1), flucytosine 1.25 microg ml(-1), fluzonazole 4 microg ml(-1), itraconazole 0.5 microg ml(-1) and ketonazole 0.25 microg ml(-1) . The agreement of the amphotericin B MIC values obtained in antibiotic medium 3 by the broth microdilution method, with those obtained on casitone medium by Etest, was within a two-dilution range for both isolates . C . laurentii may cause meningitis and may also involve the lungs in AIDS patients.

Med Mycol, 1998 Oct, 36(5), 305 - 11
Cryptococcus neoformans isolated from human dwellings in Rio de Janeiro, Brazil: an analysis of the domestic environment of AIDS patients with and without cryptococcosis; Passoni LF et al.; One hundred and fifty-four human dwellings in the metropolitan area of Rio de Janeiro, Brazil were studied . A total of 824 samples of indoor dust, outdoor soil and avian droppings were collected . Cryptococcus neoformans var . neoformans was isolated from 20 (13%) dwellings, comprising five (15.6%) of 32 dwellings of patients with AIDS-associated cryptococcosis; four (8.9%) of 45 dwellings of patients with AIDS but without cryptococcosis; and 11 (14.3%) of 77 dwellings of apparently healthy individuals (P>0.05) . The principal factor associated with domiciliar contamination by C . neoformans var . neoformans was the presence of avians in the domestic environment or nearby the home . Cryptococcosis was more frequent among AIDS patients residing in dwellings from which C . neoformans var . neoformans was isolated than among AIDS patients from whose domestic environment the fungus was not demonstrated by the methods used (odds ratio (OR)=2.05) . These findings suggest that the distribution of C . neoformans var . neoformans in Rio de Janeiro is not restricted to the classically known biotopes as well as reinforce the possibility of exogenous infection in opportunistic cryptococcosis, including exogenous infection acquired in the domestic environment.

Int J Antimicrob Agents, 1999 Jan, 11(1), 65 - 8
Characterization of antimicrobial agents extracted from Asterina pectinifera; Choi DH et al.; The extracts from the starfish Asterina pectinifera obtained using various organic solvents and distilled water were tested for their antimicrobial activities against fungi and yeasts . The methanol and water extracts were found to be the most active and Aspergillus spp . and Cryptococcus neoformans proved to be sensitive species . The extracts are polar, stable to a wide range of pH and a high temperature, and possess a very distinctive UV spectrum . The colour reactions for functional groups of antimicrobial substances indicate the presence of some distinctive chemical properties common to steroidal oligoglycosides which results suggest that the antimicrobial activity is due to structures common to steroidal oligoglycosides.

J Clin Microbiol, 1999 Apr, 37(4), 1170 - 2
Serotyping of Cryptococcus neoformans isolates from clinical and environmental sources in Spain; Baro T et al.; We determined biovars and serotypes of 154 isolates of Cryptococcus neoformans from clinical and environmental sources from different areas of Spain . All clinical isolates belonged to C . neoformans var . neoformans . Serotypes showed an irregular distribution . C . neoformans var . gattii serotype B was isolated from necropsy specimens from goats with pulmonary disease.

J Clin Microbiol, 1999 Apr, 37(4), 981 - 6
Detection of cell wall mannoprotein Mp1p in culture supernatants of Penicillium marneffei and in sera of penicilliosis patients; Cao L et al.; Mannoproteins are important and abundant structural components of fungal cell walls . The MP1 gene encodes a cell wall mannoprotein of the pathogenic fungus Penicillium marneffei . In the present study, we show that Mp1p is secreted into the cell culture supernatant at a level that can be detected by Western blotting . A sensitive enzyme-linked immunosorbent assay (ELISA) developed with antibodies against Mp1p was capable of detecting this protein from the cell culture supernatant of P . marneffei at 10(4) cells/ml . The anti-Mp1p antibody is specific since it fails to react with any protein-form lysates of Candida albicans, Histoplasma capsulatum, or Cryptococcus neoformans by Western blotting . In addition, this Mp1p antigen-based ELISA is also specific for P . marneffei since the cell culture supernatants of the other three fungi gave negative results . Finally, a clinical evaluation of sera from penicilliosis patients indicates that 17 of 26 (65%) patients are Mp1p antigen test positive . Furthermore, a Mp1p antibody test was performed with these serum specimens . The combined antibody and antigen tests for P . marneffei carry a sensitive of 88% (23 of 26), with a positive predictive value of 100% and a negative predictive value of 96% . The specificities of the tests are high since none of the 85 control sera was positive by either test.

Hunan Yi Ke Da Xue Xue Bao, 1997, 22(6), 514 - 6
{Nitric oxide, TNF-alpha and IL-8 in cerebrospinal fluids of tuberculous and cryptococcic meningitis}; Chen L et al.; The contents of NO, TNF-alpha and IL-8 in the cerebrospinal fluids (CSF) of patients with tuberculous and cryptococcic meningitis were detected . The results showed that the concentration of NO2-/NO3- and the levels of TNF-alpha and IL-8 in CSF of the two kinds of meningitis were higher than those of normal CSF, and the concentration of NO2-/NO3- correlated positively to the content of TNF-alpha . The results indicate that the over production of NO, TNF-alpha and IL-8 in CSF may involve in the tissue inflammation and damage of central nerve system infection.

J Lab Clin Med, 1999 Mar, 133(3), 274 - 88
Urokinase is required for T lymphocyte proliferation and activation in vitro; Gyetko MR et al.; We have previously demonstrated that urokinase-deficient (uPA-/-) mice do not increase lung T lymphocyte number and fail to mount protective immune responses during pulmonary Cryptococcus neoformans infection . These observations suggest a previously unconsidered role for urokinase-type plasminogen activator (uPA) in T lymphocyte-mediated immune responses . Accordingly, we sought to determine whether uPA is required for T cell receptor-mediated (TCR-mediated) lymphocyte proliferation and activation . Splenocytes from uPA-/- and uPA+/+ mice were stimulated with concanavalin A (Con A) . The uPA-/- mice had diminished T cell proliferation as compared with uPA+/+ mice . Coculturing uPA-/- T cells with uPA+/+ accessory cells led to the restoration of proliferation . Similarly, T cell proliferation induced by CD3 cross-linking was diminished in uPA-/- mice as compared with uPA+/+ mice . T lymphocyte activation, defined as the induced expression of antigens and the elaboration of cytokines, was determined . The expression of CD69 and that of CD49d were diminished in response to Con A stimulation in uPA-/- mice as compared with uPA+/+ mice . The elaboration of cytokines in response to Con A was also altered in the uPA-/- mice . The production of the Th1 cytokines interferon-gamma and interleukin-12 was diminished in uPA-/- mice as compared with uPA+/+ mice . The uPA-/- mice produced increased amounts of interleukin-10, a Th2 cytokine . We conclude that the lack of uPA results in impaired T cell activation and proliferation in response to TCR-mediated signaling and the expression of a less Th1-polarized profile of cytokines . These findings suggest that the inability of uPA-/- mice to combat Cryptococcus neoformans infection may be caused by the impairment of T lymphocyte immune responses in the absence of uPA.

Skeletal Radiol, 1999 Jan, 28(1), 49 - 51
Cryptococcoma of the sacrum; Noh HM et al.; Cryptococcoma of the sacrum was the initial presentation of systemic cryptococcosis in a patient on chronic steroid therapy for autoimmune hepatitis . The bone lesion was the only overt manifestation of systemic cryptococcal disease, which preceded other clinical manifestations and led to the subsequent diagnosis of systemic infection.

Clin Diagn Lab Immunol, 1999 Mar, 6(2), 266 - 8
Variants of a Cryptococcus neoformans strain elicit different inflammatory responses in mice; Chen LC et al.; The virulence of Cryptococcus neoformans isolates with high and low extracellular proteolytic activity was investigated in mice . No consistent relationship between proteolytic activity and virulence was observed, but isolates derived from one strain were shown to elicit different inflammatory responses.

Curr Opin Microbiol, 1998 Aug, 1(4), 381 - 9
Gene disruption to evaluate the role of fungal candidate virulence genes; Kwon-Chung K; Gene disruption is a powerful genetic tool that can define pathogenic or virulence factors . In the past two years gene disruption approaches have been used to identify fungal virulence genes . The capsule genes, an alpha subunit of G protein and certain kinases of Cryptococcus neoformans have clearly been demonstrated to be associated with pathogenicity . In Candida albicans at least four genes involved in hyphal formation have been disrupted and tested for virulence . In other fungi, such as Histoplasma capsulatum, however, more efficient gene disruption methods need to be developed before such approaches can be regularly used for identifying virulence genes.

J Zoo Wildl Med, 1998 Dec, 29(4), 474 - 8
Sporotrichosis in a nine-banded armadillo (Dasypus novemcinctus); Wenker CJ et al.; An adult female nine-banded armadillo (Dasypus novemcinctus) died in the quarantine station of a private Swiss zoo . Multifocal ulcerative skin lesions and multiple hemorrhages in the lungs were found at necropsy . The spleen was enlarged and dark red . Histologically, there was diffuse granulomatous infiltration, including multinucleated giant cells, of the skin lesions, lungs, spleen, liver, heart, and kidneys . Abundant periodic acid-Schiff-positive yeastlike cells were demonstrated intracellularly in giant cells and extracellularly scattered throughout the tissues . Morphology of the cells varied, with some nonbudding cells resembling Cryptococcus neoformans and others resembling Sporothrix schenckii . A diagnosis of sporotrichosis was confirmed by immunofluorescence studies . This is the first report of sporotrichosis in an armadillo in a zoological garden and the third report of sporotrichosis in D . novemcinctus.

No To Shinkei, 1999 Jan, 51(1), 69 - 74
{Steroid-responsive diffuse cerebral white matter lesions in a case of intractable fungal meningoencephalitis}; Suzuki Y et al.; We report a case of fungal meningoencephalitis with steroid-responsive diffuse cerebral white matter lesions . A 49-year-old male developed auditory hallucination, confusion and fever, on April, 1994 . He was diagnosed as having cryptococcal meningoencephalitis based on the detection of cryptococcal antigens in the cerebrospinal fluid (CSF) . Intravenous administration of fluconazole resulted in improvement of his neurologic symptoms and CSF findings . For the next seven months, he was treated with oral fluconazole and the neurological status was stable . However, soon after the dose of fluconazole was tapered, he became confused and febrile, which made him admitted to our hospital . Neurological examination on admission showed disturbance of consciousness, disorientation and meningeal irritation . The CSF examination revealed mild pleocytosis (mostly lymphocytes), elevated protein and normal glucose levels, although fungus was not detected . The T2-weighted image of brain MRI demonstrated diffuse hyperintense lesions in the bilateral cerebral white matters . GD-DTPA enhanced MRI showed spotty enhanced lesions in the periventricular white matters . The neurologic symptoms were once relieved after intravenous administration of fluconazole was started, but two months later, he became comatose and needed ventilatory support, despite amphotericine B therapy . Then, a needle brain biopsy targeting the white matter lesion was done . Histopathology of the specimen showed chronic inflammation with granuloma formation and T lymphycyte infiltrate around the small vessels, though fungus was not detected in the tissue . Combined therapy with corticosteroid and antifungal agents remarkably improved the neurological symptoms as well as the MRI findings . In the present case, fungal infection possibly induced an altered immune reactions which resulted in the steroid responsive diffuse cerebral white matter lesions.

Nihon Kokyuki Gakkai Zasshi, 1998 Dec, 36(12), 1038 - 42
{Pulmonary cryptococcosis exhibiting diffuse multiple nodular shadows}; Inui N et al.; We report a case of pulmonary cryptococcosis showing diffuse multiple nodular shadows in all lung fields . A 39-year-old woman with no immunological abnormalities was admitted with complaints of cough and sputum . She had experienced measles 4 weeks prior to admission . Chest x-ray films revealed diffuse nodular opacities throughout the lung fields, a finding suggestive of metastatic lung cancer . Detailed examinations, including transbronchial lung biopsy, were not conclusive . A diagnosis of pulmonary cryptococcosis was made on the basis of findings from video-assisted thoracoscopic biopsy . Primary pulmonary cryptococcosis usually appears as a solitary nodule or limited infiltration . Immunologically compromised hosts commonly demonstrate various abnormal shadows, such as the multiple nodular shadows observed in our patient . It has been reported that measles infection can cause temporary immune suppression . Secondary immunodeficiency resulting from the preceding infection with measles could explain the unusual chest x-ray findings in this case.

Eur J Immunol, 1999 Feb, 29(2), 643 - 9
Combined effects of IL-12 and IL-18 on the clinical course and local cytokine production in murine pulmonary infection with Cryptococcus neoformans; Qureshi MH et al.; We reported recently that interleukin (IL)-12 and IL-18 synergistically increased the fungicidal activity of mouse peritoneal exudate cells against Cryptococcus neoformans by inducing the production of interferon (IFN)-gamma by natural killer (NK) cells . To confirm these findings in vivo, we examined the effect of combined treatment using these two cytokines on the course of experimentally induced pulmonary and disseminated cryptococcosis in mice . IL-12 and IL-18 were used at subtherapeutic doses (0.005 and 2 microg/mouse/day, respectively) . A single administration of either cytokine was not effective in protecting mice against the infection, while combined treatment significantly prolonged survival time of infected mice and reduced the lung and brain loads of organisms . These protective effects were associated with elevated IFN-gamma and reduced IL-4 levels in bronchoalveolar lavage fluid . Finally, depletion of NK and gammadelta T cells, but not of CD4+ T cells, by administration of specific antibodies, significantly reduced the production of IFN-gamma in lungs by IL-12/IL-18 treatment during the 7 days of infection . Our results demonstrated that IL-12 and IL-18 protected mice against cryptococcal infection in a synergistic manner by enhancing the local production of IFN-gamma by NK and gammadelta T cells in the early phase of infection and by suppressing the production of IL-4 in lungs.

Clin Infect Dis, 1999 Feb, 28(2), 309 - 13
Cryptococcosis in children with AIDS; Abadi J et al.; We compiled the clinical and immunologic features of Cryptococcus neoformans infections in human immunodeficiency virus (HIV)-infected children from 1985 to 1996 in a retrospective case series . Thirty cases of cryptococcosis were identified . These children had a median age of 9.8 years, a median CD4+ cell count of 54/microL at the time of diagnosis, and either a culture positive for C . neoformans or cryptococcal antigen in serum or cerebrospinal fluid . Sixty-three percent of the cases occurred in children vertically infected with HIV and in children between 6 and 12 years of age . The clinical and laboratory characteristics of this pediatric cohort were similar to those of adults with AIDS and cryptococcosis . On the basis of a subset of the cases, a 10-year point prevalence of cryptococcosis among children with AIDS of approximately 1% was estimated.

Clin Infect Dis, 1999 Feb, 28(2), 291 - 6
A comparison of itraconazole versus fluconazole as maintenance therapy for AIDS-associated cryptococcal meningitis . National Institute of Allergy and Infectious Diseases Mycoses Study Group; Saag MS et al.; This study was designed to compare the effectiveness of fluconazole vs . itraconazole as maintenance therapy for AIDS-associated cryptococcal meningitis . HIV-infected patients who had been successfully treated (achieved negative culture of CSF) for a first episode of cryptococcal meningitis were randomized to receive fluconazole or itraconazole, both at 200 mg/d, for 12 months . The study was stopped prematurely on the recommendation of an independent Data Safety and Monitoring Board . At the time, 13 (23%) of 57 itraconazole recipients had experienced culture-positive relapse, compared with 2 relapses (4%) noted among 51 fluconazole recipients (P = .006) . The factor best associated with relapse was the patient having not received flucytosine during the initial 2 weeks of primary treatment for cryptococcal disease (relative risk = 5.88; 95% confidence interval, 1.27-27.14; P = .04) . Fluconazole remains the treatment of choice for maintenance therapy for AIDS-associated cryptococcal disease . Flucytosine may contribute to the prevention of relapse if used during the first 2 weeks of primary therapy.

J Neurol Sci, 1999 Jan 1, 162(1), 20 - 6
Meningitis in a community with a high prevalence of tuberculosis and HIV infection; Silber E et al.; OBJECTIVES: To evaluate the spectrum of aetiologies, and distinguishing clinical and laboratory features, of meningeal infection in a community with a high prevalence of tuberculosis (TB) and HIV infection . SETTING: A hospital serving mineworkers, originating from rural areas of Southern Africa . DESIGN: Prospective cohort of 60 consecutive lumbar punctures (LPs), performed for suspected meningitis . MEASUREMENTS: Clinical history and examination; concurrent cerebrospinal fluid (CSF) and blood samples; mortality status six months after entry to study . RESULTS: 38 of 57 patients (66.7%) were HIV-1 positive, 59.5% of whom had a CD4 count <200 cells/mm3 . Nine patients had tuberculous meningitis (TBM) and two had tuberculomas; four developed disease while on TB therapy . There was one case of multidrug, and two of isoniazid-resistant TBM . There were nine episodes of cryptococcal meningitis (seven patients), nine of aseptic meningitis, two of neurosyphilis and 20 normal LPs, including four with AIDS dementia complex (ADC) . Ten patients with meningococcal infection, part of a larger outbreak, were significantly younger (p=0.004) . All patients with tuberculous, cryptococcal (most immune-suppressed p<0.001) and aseptic meningitis were HIV-1 positive . Within six months, 19 patients had died . Death was associated with HIV positivity (p=0.004), low CD4 count (p<0.001) and a diagnosis of cryptococcal meningitis, CNS TB or ADC . CONCLUSION: HIV has a major impact on the burden of disease and mortality, with a predominance of opportunistic chronic meningitides, despite a meningococcal outbreak, in this community . Of concern is the development of TBM despite therapy, and the emergence of drug-resistant strains.

J Antimicrob Chemother, 1998 Dec, 42(6), 779 - 85
In-vivo therapeutic efficacy in experimental murine mycoses of a new formulation of deoxycholate-amphotericin B obtained by mild heating; Petit C et al.; Heat-induced 'superaggregation' of deoxycholate-amphotericin B (AmB-DOC, Fungizone) was shown previously to reduce the in-vitro toxicity of this antifungal agent . We compared AmB-DOC with the formulation obtained by heating the commercial form (Fungizone, Bristol Myers Squibb, Paris, France) for 20 min at 70 degrees C, in the treatment of murine infections . An improvement of antifungal activity was obtained with heated AmB-DOC formulations due to a lower toxicity which allowed the administration of higher drug doses than those achievable with the commercial preparation . Single intravenous injections of heated AmB-DOC solutions were demonstrated to be two-fold less toxic than unheated ones to healthy mice . For mice infected with Candida albicans, the maximum tolerated dose was higher with heated than with unheated AmB-DOC solutions . In the model of murine candidiasis, following a single dose of heated AmB-DOC 0.5 mg/kg, 85% of mice survived for 3 weeks, whereas at this dose the immediate toxicity of the standard formulation in infected mice restricted the therapeutic efficacy to 25% survival . Both formulations were equally effective in increasing the survival time for murine cryptococcal pneumonia and meningoencephalitis . Injection of heated AmB-DOC solutions at a dose two-fold higher than the maximal tolerated dose observed with the unheated preparation (1.2 mg/kg) increased the survival time by a factor of 1.4 in cryptococcal meningoencephalitis . These results indicate that mild heat treatment of AmB-DOC solutions could provide a simple and economical method to improve the therapeutic index of this antifungal agent by reducing its toxicity on mammalian cells.

Folia Histochem Cytobiol, 1998, 36(4), 157 - 65
The ovaries of scale insects (Hemiptera, Coccinea) . Morphology and phylogenetic conclusions; Szklarzewicz T; Coccoids (Coccinea, Coccoidea, Coccomorpha, scale insects, scales) are a highly diverse group of ectoparasitic insects . They comprise 2 subgroups: primitive archaeococcoids (= Orthezioidea sensu Koteja) and advanced neococcoids (= Coccoidea sensu Koteja) . The ovaries of coccoids consist of numerous short telotrophic-meroistic ovarioles . The ovarioles of all investigated species share common characters (e.g . the same mechanism of ovariole development, lack of terminal filaments, occurrence of single oocytes in the vitellaria) supporting the concept of monophyletic origin of this group . Despite these characteristics, the ovaries of archaeococcoids and neococcoids differ in the number of germ cells (oocytes + trophocytes) constituting a single ovariole . In primitive families (Ortheziidae, Margarodidae), this number is relatively large (15-58), whereas in advanced ones (Pseudococcidae, Kermesidae, Eriococcidae, Cryptococcidae, Coccidae, Diaspididae) it is small and usually does not exceed 8 . The comparative analysis of the ovary structure in the representatives of Coccinea and closely related Aphidinea (aphids) has revealed that: (1) the organization of archaeococcoid ovaries is more similar to those of aphids than to neococcoids and (2) during the evolution of Coccinea a gradual reduction in the number of germ cells in ovarioles took place.

Am J Respir Crit Care Med, 1999 Mar, 159(3), 733 - 40
Human immunodeficiency virus and the outcome of treatment for new and recurrent pulmonary tuberculosis in African patients; Murray J et al.; The purpose of this study was to evaluate the impact of human immunodeficiency virus (HIV) infection on treatment for tuberculosis (TB) . The study population comprised 28,522 black Southern African gold miners . Patients with sputum culture-positive new or recurrent pulmonary TB diagnosed in 1995 were prospectively enrolled in the cohort . Directly observed therapy (DOT) was practiced and outcomes were assessed at 6 mo after treatment was begun . There were 376 cases of TB (incidence 1,318 per 100,000), of which 190 (50%) were HIV positive and 82 (22%) had recurrent TB . There was no association between HIV status and history of previous TB or drug resistance . Neither the treatment interruption rate (2%) nor the rate at which patients transferred out of the treatment program (1.6%) were associated with HIV status . Excluding deaths, cure rates were similar for HIV-positive and HIV-negative patients (89% versus 88%), but significantly lower in those with recurrent than in those with new TB (77% versus 92%) . Mortality was 0.5% in HIV-negative patients versus 13.7% in HIV-positive patients, and in the latter group was associated with CD4(+) lymphocyte depletion . Autopsy examination showed that in HIV-positive patients, early mortality was due to TB whereas late deaths were most commonly due to cryptococcal pneumonia . The study showed that a well-run TB control program can result in acceptable cure rates even in a population with a very high incidence of TB and HIV infection . Particular vigilance is needed for concurrent infections, which may contribute significantly to mortality during treatment of TB in HIV-positive patients.

J Antibiot (Tokyo), 1998 Dec, 51(12), 1081 - 6
BE-31405, a new antifungal antibiotic produced by Penicillium minioluteum . I . Description of producing organism, fermentation, isolation, physico-chemical and biological properties; Okada H et al.; A new antifungal antibiotic, BE-31405, was isolated from the culture broth of a fungal strain, Penicillium minioluteum F31405 . BE-31405 was isolated by adsorption on high porous polymer resin (Diaion HP-20), followed by solvent extraction, precipitation and crystallization . BE-31405 showed potent growth inhibitory activity against pathogenic fungal strains such as Candida albicans, Candida glabrata and Cryptococcus neoformans, but did not show cytotoxic activity against mammalian cells such as P388 mouse leukemia . The mechanism studies indicated that BE-31405 inhibited the protein synthesis of C . albicans but not of mammalian cells.

Jpn J Thorac Cardiovasc Surg, 1998 Dec, 46(12), 1267 - 74
{Efficacy of F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) scans in diagnosis of pulmonary nodules}; Orino K et al.; To evaluate the value of F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) scans, we performed FDG-PET scans in 23 patients with indeterminate pulmonary nodules less than 3 cm in size and analyzed these scans qualitatively and semiquantitatively . Histologic specimens were obtained by thoracoscopic excisional biopsy in 16 patients, CT-guided needle aspiration cytology in three, and bronchoscopic brushing cytology in four . Pathological diagnoses were lung cancer in 16 patients, benign inflammation in six, and malignant lymphoma in one . Sensitivity, specificity and accuracy of the FDG-PET scans were 88% (15/17), 67% (4/6) and 83% (19/23), respectively . There were two false-positive cases (organizing pneumonia and cryptococcosis) and two false-negative ones (slow-growing adenocarcinoma and malignant lymphoma) . Although a few false-positive cases of granulomatous disease were yielded, the FDG-PET scans were highly sensitive in the detection of lung cancer . We conclude that the FDG-PET scanning in a useful diagnostic imaging modailty in the management of indeterminate pulmonary nodules.

Biochim Biophys Acta, 1999 Feb 24, 1453(2), 261 - 72
Interaction of lipopolysaccharide with human small intestinal lamina propria fibroblasts favors neutrophil migration and peripheral blood mononuclear cell adhesion by the production of proinflammatory mediators and adhesion molecules; Chakravortty D et al.; Fibroblasts are important effector cells having a potential role in augmenting the inflammatory responses in various diseases . In infantile diarrhea caused by enteropathogenic Escherichia coli (EPEC), the mechanism of inflammatory reactions at the mucosal site remains unknown . Although the potential involvement of fibroblasts in the pathogenesis of cryptococcus-induced diarrhea in pigs has been suggested, the precise role of lamina propria fibroblasts in the cellular pathogenesis of intestinal infection and inflammation caused by EPEC requires elucidation . Earlier we reported the lipopolysaccharide (LPS)-induced cell proliferation, and collagen synthesis and downregulation of nitric oxide in lamina propria fibroblasts . In this report, we present the profile of cytokines and adhesion molecules in the cultured and characterized human small intestinal lamina propria fibroblasts in relation to neutrophil migration and adhesion in response to lipopolysaccharide (LPS) extracted from EPEC 055:B5 . Upon interaction with LPS (1-10 micrograms/ml), lamina propria fibroblasts produced a high level of proinflammatory mediators, interleukin (IL)-1alpha, IL-1beta, IL-6, IL-8, tumor necrosis factor (TNF)-alpha and cell adhesion molecules (CAM) such as intercellular cell adhesion molecule (ICAM), A-CAM, N-CAM and vitronectin in a time-dependent manner . LPS induced cell-associated IL-1alpha and IL-1beta, and IL-6, IL-8 and TNF-alpha as soluble form in the supernatant . Apart from ICAM, vitronectin, A-CAM, and N-CAM proteins were strongly induced in lamina propria fibroblasts by LPS . Adhesion of PBMC to LPS-treated lamina propria fibroblasts was ICAM-dependent . LPS-induced ICAM expression in lamina propria fibroblasts was modulated by whole blood, PBMC and neutrophils . Conditioned medium of LPS-treated lamina propria fibroblasts remarkably enhanced the neutrophil migration . The migration of neutrophils was inhibited by anti-IL-8 antibody . Co-culture of fibroblasts with neutrophils using polycarbonate membrane filters exhibited time-dependent migration of neutrophils . These findings indicate that the coordinate production of proinflammatory cytokines and adhesion molecules in lamina propria fibroblasts which do not classically belong to the immune system can influence the local inflammatory reactions at the intestinal mucosal site during bacterial infections and can influence the immune cell population residing in the lamina propria.

J Gastroenterol Hepatol, 1999 Feb, 14(2), 146 - 9
Findings and benefit of liver biopsies in 46 patients infected with human immunodeficiency virus; Piratvisuth T et al.; AIMS: The aim of this work is to evaluate the role of liver biopsy and to determine the histological findings in patients infected with the human immunodeficiency virus (HIV) who have abnormal liver function tests (LFT) . METHODS: We performed a percutaneous liver biopsy in 46 HIV-seropositive patients with abnormal LFT . Parts of biopsied tissue were used for bacterial and fungal culture and the rest was processed for histological examination including special staining . RESULTS: Of these 46 patients, 41 patients were males and five were females . The median age was 31+/-6 years . Mycobacterium tuberculosis was the most common histological finding (15 cases) . Of 15 tuberculosis patients, 11 (73.3%) had lymphadenopathy and positive acid-fast bacilli (AFB) in node aspiration or biopsy . The other findings included AFB-negative granuloma (eight cases), histoplasmosis (six cases), cryptococcosis (six cases), penicillosis (four cases), viral hepatitis: hepatitis C virus (HCV; one case), hepatitis B virus and HCV infection (one case), fatty liver (two cases), drug-induced hepatitis (one case) and non-specific changes (five cases) . There were double infections in three patients . We were able to demonstrate opportunistic infections in 41 cases (89.3%) . CONCLUSIONS: Mycobacterium tuberculosis was the most common histological finding in HIV patients with abnormal LFT in Thailand . Liver biopsy was a useful procedure in evaluating abnormal LFT in HIV patients.

Clin Infect Dis, 1999 Jan, 28(1), 82 - 92
Early mycological treatment failure in AIDS-associated cryptococcal meningitis; Robinson PA et al.; Cryptococcal meningitis causes significant morbidity and mortality in persons with AIDS . Of 236 AIDS patients treated with amphotericin B plus flucytosine, 29 (12%) died within 2 weeks and 62 (26%) died before 10 weeks . Just 129 (55%) of 236 patients were alive with negative cerebrospinal fluid (CSF) cultures at 10 weeks . Multivariate analyses identified that titer of cryptococcal antigen in CSF, serum albumin level, and CD4 cell count, together with dose of amphotericin B, had the strongest joint association with failure to achieve negative CSF cultures by day 14 . Among patients with similar CSF cryptococcal antigen titers, CD4 cell counts, and serum albumin levels, the odds of failure at week 10 for those without negative CSF cultures by day 14 was five times that for those with negative CSF cultures by day 14 (odds ratio, 5.0; 95% confidence interval, 2.2-10.9) . Prognosis is dismal for patients with AIDS-related cryptococcal meningitis . Multivariate analyses identified three components that, along with initial treatment, have the strongest joint association with early outcome . Clearly, more effective initial therapy and patient management strategies that address immune function and nutritional status are needed to improve outcomes of this disease.

Am J Kidney Dis, 1999 Feb, 33(2), 282 - 6
Renal pathology and HIV infection in Thailand; Praditpornsilpa K et al.; The existence of a human immunodeficiency virus (HIV)-associated nephropathy (HIVAN) as a distinct disease entity characterized by glomerulosclerosis is well established in North America and Western Europe . Although the large number of HIV-infected cases overwhelm the Asian countries, no cases of HIVAN are documented in the literature . We studied 26 cases of HIV-infected Thai patients with proteinuria greater than 1.5 g/d of protein during 1995 and 1996 . None of the patients were treated with antiretroviral drugs at the time of renal biopsy . Intravenous drug addiction and sexual transmission were risk factors in 11 and 15 patients, respectively . Pathological examinations were performed by light microscopic and immunoperoxidase study . Mesangial proliferative glomerulonephritis was found in 17 cases, immunoglobulin A (IgA) nephropathy in 2 cases, and diffuse proliferative glomerulonephritis and interstitial nephritis secondary to cryptococcal infection in 2 cases each . One case each had membranous glomerulopathy, membranoproliferative glomerulonephritis, and granulomatous interstitial nephritis secondary to tuberculosis . The renal pathological findings of HIVAN with the unique features described in previous literature were not evident in these patients . Although the data in this study are limited to 26 HIV-infected Thai patients, we believe that HIVAN is uncommon in the Asian HIV-infected population.

Gene, 1999 Feb 18, 227(2), 231 - 40
The yeast Cryptococcus neoformans uses 'mammalian' enhancer sites in the regulation of the virulence gene, CNLAC1; Zhang S et al.; Transcriptional regulation in mammalian and plant cells is distinguished from fungi by the presence of blocks of multiple interacting DNA binding sites distributed over a relatively large upstream region of genes and the ability to use glutamine-rich enhancers such as Sp1 . We offer evidence that the haploid yeast Cryptococcus neoformans contains a virulence gene, CNLAC1, having regulatory properties more similar to mammalian systems than to that of yeast . We used a novel promoter plasmid, pVEW, and electromobility shift assay techniques adapted for the fungus for the first systematic structural and functional study of a 5'-enhancer region of a basidiomycete fungus using the upstream region of CNLAC1 . Two groups of interactive enhancer regions, located over a range of 1.5kb from the mRNA start site are involved in CNLAC1 regulation (region 2: -1721 to -1615 and region 7) in addition to a TATA promoter at position -539 . Region 2 contains a consensus Sp1 site and region 7 contains a consensus E2F site, each of which shows significant binding to nuclear proteins under derepressed conditions; cooperative binding was also suggested between DNA-binding protein of these sites and those binding nearby CCAAT sequences in each region . Two regions of repression were also evident under derepressed conditions (region 5: -1351 to -1207 and region 8: -991 to -971) . Identification of functional Sp1 binding sites and the presence of multiple interactive enhancer sites over a fairly large upstream range suggests that cryptococcal transcriptional regulation contains features often associated with higher eukaryotic regulation . C . neoformans thus may provide a unique system for the study of certain aspects of higher eukaryotic transcription, using yeast genetic approaches . In addition, properties of basidiomycete yeast such as Cryptococcus exemplified in the present study suggest an evolutionary progression in gene regulation within fungi toward properties exhibited in the kingdoms Animalia and Plantae.

Med Mycol, 1998, 36 Suppl 1, 57 - 67
Development of vaccines and their use in the prevention of fungal infections; Dixon DM et al.; Vaccine approaches to infectious diseases are widely applied and appreciated . Disciplines such as bacteriology and virology have a rich history of successful vaccine development . The complexity of eukaryotic systems presents additional challenges to the development of vaccines against them . These challenges are being met in the fields of parasitology, and are being revisited for application in oncology . Vaccine opportunities exist in medical mycology . The National Institute of Allergy and Infectious Diseases has held a series of workshops in medical mycology where the need to develop vaccines for fungal diseases was noted and where important opportunities were discussed . Major advances in vaccinology and the technology of antigen preparation and delivery have increased feasibility and heightened interest . The recent epidemic of coccidioidomycosis in the American Southwest has demonstrated the need for developing a vaccine as an effective preventive measure for those living in and for those who subsequently move into regions with the endemic mycoses . The XIIth Congress of the International Society for Human and Animal Mycology included a symposium that summarized new vaccination strategies for selected fungi: Candida albicans, Coccidioides immitis, and Trichophyton verrucosum . The goal of the present summary is to provide representative examples of continuing efforts relating to vaccine development within the medical mycological community highlighting Blastomyces dermatidis, Cryptococcus neoformans, Histoplasma capsulatum, Paracoccidioides brasiliensis, and Pythiumn insidiosum.

Med Mycol, 1998, 36 Suppl 1, 249 - 57
Molecular diagnosis and epidemiology of fungal infections; Reiss E et al.; A variety of methods are utilized for DNA strain subtyping of Candida spp . because no 'gold standard' exists . Random amplified polymorphic DNA (RAPD) or restriction enzyme analysis (REA) are useful to determine the source of an outbreak, but more reproducible and discriminatory methods such as Southern hybridization and pulsed field gel electrophoresis (PFGE) may be required . When applied to some nosocomial Candida infections, multiple strains and species have been identified . Microevolution of yeast species occurs and epidemiologically related isolates may show minor pattern differences, creating uncertainty as to whether they are distinct strains . Approximately 1000 isolates of Aspergillus fumigatus from environmental and clinical sources were typed by REA probed with an A . fumigatus-specific retrotransposon-like sequence . Patients with no symptom of aspergillosis may carry several strains, whereas patients with pulmonary aspergillosis may carry one or two strains; nocosomial transmission of aspergillosis was proven in 39% of the patients studied; any given environmental strain can be infectious; the environmental population of A . fumigatus is extremely diverse and no specific niche was found in the hospital . A PCR assay was designed to target conserved 18S-ribosomal DNA (rDNA) sequences shared by most fungi and a 687 bp product was amplified from 25 medically important fungal species . Studies with blood, cerebrospinal fluid and sputum specimens from patients with mycoses indicated that the PCR assay is more sensitive in diagnosing invasive fungal infections than blood culture methods . More specific identification is obtainable with genus/species-specif c probes designed from within the PCR-amplified sequences for C . albicans, C . krusei, C . lusitaniae, Pneumocystis carinii, Cryptococcus neoformans, Aspergillus/Penicillium spp . and C . glabrata/Saccharomyces cerevisiae . A . fumigatus and A . niger were differentiated by denaturing gradient gel electrophoresis . In situ hybridization (ISH) detected a 648 bp fragment of the 18S rDNA of C . neoformans and a 568 bp fragment of the alkaline proteinase gene of A . fumigatus in tissues from experimentally infected animals . In ISH, the entire process can be automated, making this procedure rapid and easy . The difficulty in establishing a diagnosis of invasive candidiasis has prompted the quest for a clinically useful PCR test for candidaemia . The universal fungal oligonucleotide primer pair, ITS3 and ITS4, amplifies portions of the 5.8S ad 28S rDNA subunits, and the ITS2 region . Although rRNA genes are highly conserved, the ITS regions are distinctive . DNA probes were designed from ITS2 that were specific for 16 different Candida species . Simple, rapid sample preparation was suitable for PCR analysis of BacT/Alert blood culture bottles . Sample preparation, PCR, and EIA detection of the amplicon from five different Candida species was accomplished in 7 h, 2.5 days sooner than by conventional culture methods . As well as saving time, minor yeast species among a major species, or among bacteria, were simultaneously detected . PCR-EIA using a microtitration plate format had sensitivity 10-times greater than that obtained with ethidium bromide-stained agarose gels . Taqman combines in one step PCR, probe hybridization, and fluorescent signal generation . Taqman PCR had sensitivity equivalent to PCR-EIA and required only 5 h, including sample preparation.

Med Mycol, 1998, 36 Suppl 1, 207 - 15
Recent advances in cryptococcosis, candidiasis and coccidioidomycosis complicating HIV infection; Kappe R et al.; Concomitant with the decline in CD4+ T-cells seen as human immunodeficiency virus (HIV) infection progresses, the prevalence of opportunistic mycoses increases dramatically . This article reviews selected recent advances in our understanding of the immunology, molecular epidemiology and treatment of fungal infections in patients infected with HIV . For cryptococcosis, studies are reported on how HIV infection affects the immune response to Cryptococcus neoformans and, conversely, how stimulation with C . neoformans induces HIV production from latently HIV-infected cells . In addition, studies are presented examining the efficacy of triple combination antimycotic chemotherapy in cryptococcosis . For candidosis, investigations into genetic profiles of Candida albicans isolates obtained from patients, with resistance to antifungal agents, are demonstrated . Finally, for coccidioidomycosis, prospective studies are presented examining the clinical, epidemiological and immunological characteristics of a cohort of HIV-infected subjects residing in an endemic area.

Med Mycol, 1998, 36 Suppl 1, 119 - 28
Antifungal drug resistance in pathogenic fungi; Vanden Bossche H et al.; Failures of drug treatment in fungal infections combined with improvements in performances and standardization of antifungal susceptibility testing have drawn attention to the problem of antifungal resistance and its underlying mechanisms . Resistance of Candida species and Cryptococcus neoformans to flucytosine (5FC) develops during monotherapy . Acquired resistance results from a failure to metabolize 5FC to 5FUTP and 5FdUMP, or from the loss of feedback control of pyrimidine biosynthesis . A combination of 5FC and amphotericin B (AmB) reduces the appearance of resistant C . albicans isolates . Resistance to AmB is unusual . C . lusitaniae is the most susceptible to AmB resistance . C . neoformans with decreased AmB susceptibility has been isolated from an HIV-infected patient . Acquired resistance to AmB is often associated with alteration of membrane lipids, especially ergosterol . Concomitant with the widespread use of fluconazole there have been increasing reports of fluconazole resistance in Candida species and C . neoformans . Fluconazole resistance was mostly associated with prior use of fluconazole as intermittent therapy or prophylactic continuous treatment for recurrent thrush . In contrast to fluconazole, itraconazole is active against C . krusei . Decreased susceptibility to itraconazole is observed over time in C . albicans isolates becoming resistant to fluconazole . Decreased susceptibility to itraconazole and SCH-56592 was also observed in a few Aspergillus fumigatus isolates . Failure to accumulate azole antifungals has been identified as a cause of resistance in several post-treatment C . albicans, C . glabrata and C . krusei isolates . In azole-resistant C . albicans isolates from AIDS patients with oropharyngeal candidiasis, multidrug efflux transporters of the ATP-binding cassette (ABC) superfamily and of the class of major facilitators (MF) have been shown to be responsible for the low level of accumulation of azole antifungal agents . Two genes for these transporters, the ABC-transporter gene CDR1 and the MF gene, CaMDR1 (BEN) were shown to be overexpressed in resistant C . albicans isolates . Overexpression of BEN in Saccharomyces cerevisiae conferred resistance to fluconazole and terbinafine . CDR1 overexpression in S . cerevisiae conferred cross-resistance to fluconazole, itraconazole, ketoconazole and terbinafine . C . albicans clinical isolates resistant to azole antifungal agents over-expressing the ABC-transporter genes CDR1 and CDR2 were less susceptible to the morpholine derivative amorolfine . In C . glabrata isolates azole resistance is based on over-expression of the CgCDR gene . A reduced susceptibility of ergosterol biosynthesis is another mechanism of resistance described in a number of post-treatment C . albicans, C . neoformans and Histoplasma capsulatum isolates . Mutations have been reported in the CYP51A1 genes of resistant C . albicans isolates . Over-expression of CYP51A1 in C . albicans and C . glabrata may also account for a decreased susceptibility to azole antifungal agents.

Med Mycol, 1998, 36 Suppl 1, 79 - 86
Cryptococcus neoformans: virulence and host defences; Perfect JR et al.; Cryptococcus neoformans represents a model organism for the study of virulence and the host response . In this discussion, there is a focus on the genetic, molecular, and biochemical aspects of C . neoformans as it interacts with the host . Investigations into direct and indirect virulence phenotypes are now possible . The molecular aspects of two major virulence factors, capsule and melanin, are characterized . Yeast polyol metabolism through mannitol is examined as a potential biochemical pathway for virulence . The concept of C . neoformans differentially expressed genes within the host or in response to certain environmental cues can be used indirectly to identify potential virulence genes . However, despite significant progress in molecular pathogenesis with C . neoformans, the future of research in this area will require a certain critical mass of investigators to help share in the developmental costs which continue to occur.

Med Mycol, 1998, 36 Suppl 1, 38 - 44
Fate of transforming DNA in pathogenic fungi; Kwon-Chung KJ et al.; Genetic engineering is an important tool in helping us to define the molecular basis of pathogenicity and is also useful in helping us to identify new therapeutic targets in pathogenic fungi . Molecular genetic manipulation of micro-organisms requires the development of plasmid-mediated transformation systems that include: (i) infusion of exogenous DNA into recipient cells, (ii) expression of genes present on the incoming DNA, and (iii) stable maintenance and replication of the inserted DNA leading to expression of the desired phenotypic trait . Transformation systems have been developed for only a handful of fungi that are pathogenic to humans including several species of Candida, Cryptococcus neoformans, Histoplasma capsulatum, Blastomyces derrmatitidis, Aspergillus fumigatus, Wangiella dermatitidis (Exophiala dermatitidis) and Coccidioides immitis . Except for Candida species and A . fumigatus, where passage of exogenous DNA into recipient cells has been achieved readily using methods developed for transformation of Saccharomyces cerevisiae and Aspergillus nidulans, respectively, development of transformation systems in other pathogenic fungi has been delayed considerably and has only been possible recently with the introduction of electroporation and biolistic methods . Conventional spheroplasting methods or cell wall permeabilization methods using lithium acetate have not been successful for transformation of C . neoformans and work with only low efficiency in H . capsulatum . The fate of incoming DNA varies greatly in these pathogenic species regardless of their phylogenetic relationships . Understanding the fate of incoming DNA is critical for the construction of transforming vectors and the molecular manipulation of the organisms . In this symposium, recent advances in molecular genetic systems including transformation systems, the fate of incoming DNA and strategies for targeted integration are discussed in relation to four pathogenic fungi.

Mayo Clin Proc, 1999 Jan, 74(1), 78 - 100
Antifungal agents . Part II . The azoles; Terrell CL; Before 1978, amphotericin B and flucytosine were the only drugs available for the treatment of systemic fungal infections . The imidazoles, miconazole and ketoconazole, were introduced during the next 3 years . Intravenously administered miconazole served a limited therapeutic role and is no longer available . Orally administered ketoconazole, an inexpensive, effective, and convenient option for treating mucosal candidiasis, was widely used for a decade because it was the only available oral therapy for systemic fungal infections . During the 1990s, use of ketoconazole diminished because of the release of the triazoles--fluconazole and itraconazole . Fluconazole is less toxic and has several pharmacologic advantages over ketoconazole, including penetration into the cerebrospinal fluid . In addition, it has superior efficacy against systemic candidiasis, cryptococcosis, and coccidioidomycosis . Despite a myriad of drug interactions and less favorable pharmacologic and toxicity profiles in comparison with fluconazole, itraconazole has become a valuable addition to the antifungal armamentarium . It has excellent activity against sporotrichosis and seems promising in the treatment of aspergillosis . Itraconazole has replaced ketoconazole as the therapy of choice for nonmeningeal, non-life-threatening cases of histoplasmosis, blastomycosis, and paracoccidioidomycosis and is effective in patients with cryptococcosis and coccidioidomycosis, including those with meningitis . Further investigation into the development of new antifungal agents is ongoing.

J Clin Microbiol, 1999 Mar, 37(3), 838 - 40
Cryptococcus neoformans var . grubii: separate varietal status for Cryptococcus neoformans serotype A isolates; Franzot SP et al.; Cryptococcus neoformans var . neoformans presently includes isolates which have been determined by the immunologic reactivity of their capsular polysaccharides to be serotype A and those which have been determined to be serotype D . However, recent analyses of the URA5 sequences and DNA fingerprinting patterns suggest significant genetic differences between the two serotypes . Therefore, we propose to recognize these genotypic distinctions, as well as previously reported phenotypic differences, by restricting C . neoformans var . neoformans to isolates which are serotype D and describing a new variety, C . neoformans var . grubii, for serotype A isolates.

J Clin Microbiol, 1999 Mar, 37(3), 715 - 20
Genetic multilocus studies of different strains of Cryptococcus neoformans: taxonomy and genetic structure; Bertout S et al.; The genotypes of 107 strains of Cryptococcus isolated from the environment or from patients from various geographical areas were determined by multilocus enzyme electrophoresis (MLEE) . We analyzed the relationships between genotype structure and serotype and between genotype structure and strain origin . Twelve of the 14 enzyme-encoding loci studied were polymorphic, giving rise to 48 electrophoretic types . The genotypes of C . neoformans and C . laurentii were very similar . MLEE could not distinguish between these two pathogenic species . A correlation between the genetic multilocus structure and the origin of the sample (from the environment or patients) existed . A second analysis detected a correlation between genotype distribution and serotype . The second analysis considered three serotype groups (B, C, and A plus D plus A/D), proving that serotypes A, D, and A/D are closely related . MLEE is a useful epidemiological tool for improving our understanding of the biology of this fungus.

J Clin Microbiol, 1999 Mar, 37(3), 694 - 9
PCR detection of DNA specific for Trichosporon species in serum of patients with disseminated trichosporonosis; Nagai H et al.; Deep-seated trichosporonosis is a lethal opportunistic infection that disseminates rapidly and widely in immunocompromised patients, and early diagnosis is crucial for the treatment of this infection . We developed a novel nested-PCR assay that detects DNA specific for clinically important strains of Trichosporon in serum samples from patients with disseminated trichosporonosis . In this assay, two sets of oligonucleotide primers were derived from the sequence of 26S rRNA genes of Trichosporon asahii . The specific fragment was amplified from T . asahii and T . mucoides, but not from other microorganisms, including some other basidiomycetous fungi (Cryptococcus, Malassezia, Rhodotorula, and Sporobolomyces) . Target DNA was detected by the nested PCR with as little as 5 fg of the extracted DNA of T . asahii . In a study using 11 clinical samples, the specific fragment was detected by the nested PCR in 64% (7 of 11) of sera from patients with histologically diagnosed disseminated trichosporonosis, while glucuronoxylomannan antigen was detected in only 54% (6 of 11) of the samples . Our new nested-PCR assay using serum samples can be performed repeatedly throughout the course of the disease . In addition, not only can it be used for early diagnosis of trichosporonosis, but it may also be beneficial for monitoring its progress or response to therapy.

J Clin Microbiol, 1999 Mar, 37(3), 591 - 5
Multicenter comparison of the sensititre YeastOne Colorimetric Antifungal Panel with the National Committee for Clinical Laboratory standards M27-A reference method for testing clinical isolates of common and emerging Candida spp., Cryptococcus spp., and other yeasts and yeast-like organisms; Espinel-Ingroff A et al.; National Committee for Clinical Laboratory Standards (NCCLS) standard guidelines are available for the antifungal susceptibility testing of common Candida spp . and Cryptococcus neoformans, but NCCLS methods may not be the most efficient and convenient procedures for use in the clinical laboratory . MICs of amphotericin B, fluconazole, flucytosine, itraconazole, and ketoconazole were determined by the commercially prepared Sensititre YeastOne Colorimetric Antifungal Panel and by the NCCLS M27-A broth microdilution method for 1,176 clinical isolates of yeasts and yeast-like organisms, including Blastoschizomyces capitatus, Cryptococcus spp., 14 common and emerging species of Candida, Hansenula anomala, Rhodotorula spp., Saccharomyces cerevisiae, Sporobolomyces salmonicolor, and Trichosporon beigelii . Colorimetric MICs of amphotericin B corresponded to the first blue well (no growth), and MICs of the other agents corresponded to the first purple or blue well . Three comparisons of MIC pairs by the two methods were evaluated to obtain percentages of agreement: 24- and 48-h MICs and 24-h colorimetric versus 48-h reference MICs . The best performance of the YeastOne panel was with 24-h MICs (92 to 100%) with the azoles and flucytosine for all the species tested, with the exception of C . albicans (87 to 90%) . For amphotericin B, the best agreement between the methods was with 48-h MIC pairs (92 to 99%) for most of the species tested . The exception was for isolates of C . neoformans (76%) . These data suggest the potential value of the YeastOne panel for use in the clinical laboratory.

J Immunol, 1999 Feb 1, 162(3), 1618 - 23
Specific activated T cells regulate IL-12 production by human monocytes stimulated with Cryptococcus neoformans; Retini C et al.; IL-12 production mediated by a T cell-independent and/or T cell-dependent pathway was investigated in human monocytes responding to Cryptococcus neoformans . The data of this study showed that: 1) appreciable levels of IL-12 were observed when freshly isolated monocytes were exposed to acapsular C . neoformans or Candida albicans and secretion occurred within 24-48 h of incubation; 2) monocytes alone were poor producers of IL-12 when stimulated with encapsulated C . neoformans; 3) the presence of specific anti-glucuronoxylomannan mAb favored IL-12 secretion and Fc cross-linking could play a role; 4) monocytes were able to secrete consistent levels of IL-12 when cultured with activated T cells responding to C . neoformans; 5) the maximum secretion of IL-12 was observed at 5-7 days of culture and was strongly regulated by the presence of endogenous IFN-gamma; and 6) the interaction between CD40 on monocytes and CD40 ligand on activated T lymphocytes responding to C . neoformans played a critical role in IL-12 secretion . These data highlight the mechanisms of IL-12 production by human monocytes exposed to C . neoformans, indicating a possible biphasic secretion of IL-12, dependent on the direct effect of fungal insult, and characterized by consistent secretion of IL-12 that is dependent on the interaction of CD40 with the CD40 ligand expressed on activated T cells responding to C . neoformans.

Recenti Prog Med, 1998 Dec, 89(12), 657 - 67
{AIDS and opportunistic visceral mycoses . A diagnostic and therapeutic update}; Manfredi R; Candidiasis, cryptococcosis, aspergillosis, histoplasmosis, and coccidioidomycosis are the most commonly recognized visceral mycoses complicating the course of HIV infection and AIDS . Their current management issues are discussed on the ground of the personal experience, and the most recent literature evidences.

Nippon Ishinkin Gakkai Zasshi, 1999, 40(1), 31 - 4
Ploidy of serotype AD strains of Cryptococcus neoformans; Tanaka R et al.; Twenty-four serotype AD strains of Cryptococcus neoformans were tested for ploidy and mating type . These included 13 natural isolates (1 from a patient, 1 from peach juice, 11 from pigeon droppings), 2 single clones from one of these natural isolates, 7 F1 progeny from a self-fertile strain and 2 F1 progeny of a cross between a strain from a patient (serotype A, alfa-mating type: MATalfa) and a tester strain (serotype D, a-mating type: MATa) . Six strains (2 of natural isolates, 1 of single clone and 3 of F1 progeny) were MATalfa, 1 of F1 progeny was MATa, 4 (1 of natural isolate, 1 of single clone and 2 of F1 progeny) were a-alfa-mating type (MATa-alfa) and 13 (10 of natural isolates and 3 of F1 progeny) were untypable . Most strains, regardless of mating type, were diploid.

Nippon Ishinkin Gakkai Zasshi, 1999, 40(1), 9 - 14
{Dermatomycosis in human and animals}; Nakamura Y et al.; Dermatomycosis including dermatophytosis, sporotrichosis and cryptococcosis commonly occurs in humans and animals all and are considered to be zoonotic diseases . Recently, human cases of dermatophytosis transmitted from animals are increasing in number due to changes in the environments of human and animal life . Three species of dermatophytes, Microsporum canis, Trichophyton mentagrophytes and T . verrucosum are the most important pathogens from animal to human, respectively . Therefore, it is necessary to understand their biological and ecological characteristics to correctly diagnose and treat the disease . Some human cases of sporotrichosis and cryptococcosis were reported to be transmitted from animals in Europe and America, suggesting that medical doctors should be careful in taking the history of human patients with reference to their contacts with animals . Close cooperation between medical and veterinary doctors is required in clinical studies on mycotic infection.

Mycopathologia, 1998, 142(2), 71 - 6
Comparison of phospholipase production in Cryptococcus neoformans isolates from AIDS patients and bird droppings; Vidotto V et al.; Secreted phospholipase has been recently proposed as a virulence determinant in Cryptococcus neoformans as well as Candida albicans . This issue of cryptococcal phospholipase requires screening of phospholipase production in a larger number of isolates from clinical and environmental sources . In this study we examined phospholipase production in a total of 67 C . neoformans isolates from AIDS patients and bird droppings by using the egg-yolk plate method . Phenoloxidase activity, capsule size and growth at 37 degrees C were also measured in these strains in order to observe a possible relationship between phospholipase production of different C . neoformans strains and its virulence . Four of the 21 AIDS strains at 28 degrees C and 1 at 37 degrees C did not produce phospholipase, respectively . In contrast, 38 and 34 of the 46 bird dropping strains were negative for phospholipase production at 28, and 37 degrees C, respectively . Statistical analysis revealed a significant difference in phospholipase production, capsule size and growth ability at 37 degrees C, but not phenoloxidase activity, between the AIDS and the bird dropping strains . The highly prevalent distribution of phospholipase activity in the AIDS strains suggests a role of the enzyme in invading the host.

Antimicrob Agents Chemother, 1999 Feb, 43(2), 413 - 4
An alternative animal model for comparison of treatments for cryptococcal meningitis; Najvar LK et al.; Weanling outbred rats were infected with Cryptococcus neoformans by direct percranial puncture and inoculation into the cranium . A lethal infection ensued . Treatment with LY295337, a depsipeptide with antifungal activity, was effective in prolonging survival and reducing fungal counts in brain tissue . Weanling rats are an acceptable model for the study of central nervous system infection with C . neoformans.

Antimicrob Agents Chemother, 1999 Feb, 43(2), 233 - 9
Amphotericin B and fluconazole affect cellular charge, macrophage phagocytosis, and cellular morphology of Cryptococcus neoformans at subinhibitory concentrations; Nosanchuk JD et al.; Amphotericin B (AmB) and fluconazole (FLU) are the major antifungal drugs used in the treatment of cryptococcosis . Both drugs are believed to exert their antifungal effects through actions on cell membrane sterols . In this study we investigated whether AmB and FLU had other, more subtle effects on C . neoformans that could contribute to their therapeutic efficacy . C . neoformans cells were grown in media with subinhibitory concentrations of either AmB or FLU and analyzed for cellular charge, phagocytosis by macrophages with antibody and complement opsonins, appearance by scanning electron and light microscopies, and release of the capsular polysaccharide glucuronoxylomannan into the culture medium . Growth in the presence of either AmB or FLU resulted in major reductions in cellular charge, as measured by determination of the zeta potential . Phagocytosis studies demonstrated that exposure of C . neoformans to subinhibitory concentrations of AmB or FLU enhanced phagocytosis by macrophages . Scanning electron microscopy revealed that a large proportion of cells had an altered capsular appearance . Cells grown in medium with either AmB or FLU were smaller and released more glucuronoxylomannan into the culture medium than cells grown without antibiotics . The results suggest additional mechanisms of action for AmB and FLU that may be operative in body compartments where drug levels do not achieve the MICs . Furthermore, the results suggest mechanisms by which AmB and FLU can cooperate with humoral and cellular immune defense systems in controlling C . neoformans infections.

Med Clin (Barc), 1998 Dec 5, 111(19), 725 - 30
{Pulmonary cavitation lesions in patients infected with the human immunodeficiency virus: an analysis of a series of 78 cases}; Rodriguez Arrondo F et al.; BACKGROUND: To assess the clinical, radiologic and microbiological features of lung cavitation and HIV infection . Evaluation of the differences related to this disease in the last years . PATIENTS AND METHODS: Retrospective review of all patients with lung cavitation and HIV infection admitted at our hospital from January 1989 until December 1994 and prospective study of all patients with the same characteristics during 1995 and 1996 . Lung cavitation was defined as any parenchymal lesion, with air content, visible in a simple X-ray and greater than 1 cm of diameter . Criteria for confirmed, probable or possible diagnosis were defined . RESULTS: 78 cases of lung cavitation have been identified in 73 patients . The radiologic patterns included unilobar and multilobular involvement in 31 and 47 cases, respectively . Cavities were multiple and single in 40 and 38 cases respectively . Findings with fine needle aspiration biopsy (FNAB) were diagnostic in 11 out of 14 cases . A clinical diagnosis was performed in all 78 cases, with microbiological results in 69 cases (88.5%): Mycobacterium tuberculosis in 20, Pneumocystis carinii in nine, Pseudomonas aeruginosa in nine, Staphylococcus aureus in eight (5 endocarditis with cavitary septic emboli), Rhodococcus equi in six, P . aeruginosa and S . aureus in three, Salmonella enteritidis in three, Cryptococcus neoformans in two, Aspergillus fumigatus in two and others in 7 cases . Confirmed, probable and possible diagnosis was considered in 54, 15 and 9 cases, respectively . Thirteen episodes of spontaneous pneumothorax were found . CONCLUSIONS: The lung cavitation rate is low, compared with the number of admissions related to HIV infection; nevertheless, many of them are in close relationship with HIV infection, and most of them are caused by treatable infections . It is important to know the clinical and radiological characteristics, in order to establish an early diagnosis and an appropriate therapy . Pseudomonas aeruginosa is becoming an important cause of lung cavitation . In our series, spontaneous pneumo-thorax was not related to Pneumocystis carinii pneumonia in 61.5% of cases.

Rev Hosp Clin Fac Med Sao Paulo, 1998 Jul-Aug, 53(4), 184 - 8
{Immunohistochemistry diagnosis of fungal infections}; Rocha DC et al.; Because the mycosis incidence has increased a lot with the appearing of AIDS, the Immunohistochemistry study among fungus shows the importance of fast methods for their identification that have advantage of been a durable method in comparison with immunofluorescence and the possibility of making retrospective studies in material embedded in paraffin . The Immunohistochemistry reaction with Histoplasma capsulatum, Pneumocystis carinii; and Criptococcus neoformans antibodies were sensitive, specifics, and intensely positive in all the cases previously diagnosed as Histoplasmosis, Pneumocystosis and Cryptococcosis, without cross-reaction with other fungus; while the anti-Candida albicans antibody showed weak positiveness in four Histoplasmosis cases, in one of Paracoccidioidomycosis cases and Sporotrichosis case; and the reactions with the antibody anti-P . brasiliensis were intensely positive in all the Paracoccidioidomycosis cases and weakly positive in two Histoplasmosis and two of the four Candidiasis cases . The previous identification of each fungi on tissue sample was made by Grocott method . This preliminary study showed that it is necessary to use other kinds of antibody and fungus, in order to get more details about the possible occurrence of cross-reactions . We suggest the use of new antibodies, with new standardizations in order to find the best titles for each reaction and eliminate the cross-reactions.

J Thorac Imaging, 1999 Jan, 14(1), 51 - 62
Opportunistic fungal pneumonia; Connolly JE Jr et al.; Opportunistic fungal infection is a common cause of serious morbidity and mortality in immunocompromised patients . These infections occur primarily in patients with chemotherapy-induced neutropenia, acquired immunodeficiency syndrome . or immunosuppression after solid organ or bone marrow transplantation . The most important opportunistic fungal pathogens include Cryptococcus neoformans, Candida and Aspergillus species, and the fungi that cause mucormycosis . Opportunistic pneumonia caused by previously unrecognized pathogens, such as Fusarium, Penicillium, and the dematiaceous fungi, are increasingly reported . The clinical and radiologic features of opportunistic fungal pneumonia are highly variable and often nonspecific . Diagnosis requires knowledge of the various modes of presentation, radiologic manifestations, and epidemiology of these infections . Because many of these organisms can colonize the upper airway, sputum cultures are considered diagnostically unreliable . Instead, definitive diagnosis requires culture of the fungus from infected tissue or demonstration of the organism on microscopic examination.

Am J Pathol, 1999 Jan, 154(1), 145 - 52
Expression of inducible nitric oxide synthase in human granulomas and histiocytic reactions; Facchetti F et al.; Inducible nitric oxide synthase (iNOS) is required in immune response against infections and is involved in granuloma formation in animals; in murine macrophages, iNOS is induced by lipopolysaccharide and interferon-gamma . In contrast, the role of iNOS in human immune response against infections is still questioned, and its expression in granulomas is poorly investigated . Using Western blotting and immunohistochemistry, we investigated iNOS expression in human lymph nodes with nonspecific reactions and in tissues containing granulomas caused by mycobacteria, Toxoplasma, Cryptococcus neoformans, Leishmania, Bartonella, noninfectious granulomas (sarcoidosis, foreign body), and other hystiocitic reactions (Kikuchi's disease, Omenn syndrome) . iNOS was undetectable in nonspecific reactive lymphadenitis, foreign-body granulomas, and Omenn syndrome, whereas it was strongly expressed in infectious granulomas, sarcoidosis, and Kikuchi's diseases . Immunohistochemistry demonstrated that iNOS was selectively expressed by the epithelioid and multinucleated giant cells within the granulomas . Use of an anti-nitrotyrosine antibody, recognizing nitrosilated amino acid residues derived from nitric oxide production, revealed a consistent positivity within the cells expressing iNOS, thus suggesting that iNOS is functionally active . Detection of cytokines by reverse transcriptase-polymerase chain reaction demonstrated that tissues that were positive for iNOS, also expressed the Thl-type cytokine interferon-gamma mRNA, but not the Th2-type cytokine interleukin-4 . Taken together, these results indicate that iNOS is involved in different human immune reactions characterized by histiocytic/granulomatous inflammation and associated with Th1-type cytokine secretion.

Mycoses, 1998 Nov, 41(9-10), 389 - 96
Phenotypic and genotypic differentiation of several human and avian isolates of Cryptococcus neoformans; Hotzel H et al.; The Cryptococcus neoformans strains isolated from two human cases could be diagnosed as Cr . neoformans var . neoformans by differentiation on the basis of their characteristics determined by proline, canavanine and EDTA urease tests . The results of the serovar assignment were: for the isolate from the meningoencephalitis patient with lethal outcome, serovar A; for the strain isolated from the osteomyelitis patient with benign course, serovar D . Also, the PCR fingerprinting using primers (GACA)4, (CAC)5 and FM 1 resulted in a clear and reproducible assignment of the Cr . neoformans strains to the varieties neoformans and gattii, respectively, and, in addition, it confirmed the serovar assignment . No statistically confirmed differences in virulence between the osteomyelitis and the meningoencephalitis strain could be established by i.v . testing in mice, nor did the PCR with several primers provide any clues to a genetically determined higher virulence of the meningoencephalitis strain . The different classification as serovars A and D does not allow any conclusions concerning different virulence . It was not possible to retrospectively establish the sources of infection of the two Cr . neoformans infections, but pigeon faeces may well have played a role as a reservoir for one of the illnesses.

Kansenshogaku Zasshi, 1998 Dec, 72(12), 1261 - 8
{Clinical study on fluconazole (FLCZ) in the treatment of primary pulmonary cryptococcosis}; Hayashi Y et al.; We treated two cases of primary pulmonary cryptococcosis with fluconazole (FLCZ), the clinical usefulness of FLCZ was evaluated . FLCZ was administered orally in doses of 300 mg daily for about six months . Concentrations of FLCZ were measured in the serum of the two cases and in the bronchoalveolar lavage (BAL) fluid in one case . The following results were obtained: 1 . Clinical cures were obtained in the two cases . 2 . The serum levels of FLCZ was 15.1 microliters/ml, 13.6 micrograms/ml two hours after administration of 100 mg in case 1, that of levels were 11.1 micrograms/ml, 8.9 micrograms/ml one hour and 4.5 hours, respectively, after administration of 100 mg in case 2 . BAL was performed 4.5 hours after administration of 100 mg in case 2, the BAL fluid level of FLCZ was 0.7 microgram/ml . 3 . The minimal inhibitory concentration of FLCZ against one strain obtained from the cytology brush in case 1 was 4.0 micrograms/ml . 4 . The cryptococcal antigen titer decreased with the improvement of clinical signs and the resolution of chest X-ray abnormalities within about six months, and there was no relapse . From these results, we consider that FLCZ is a useful antifungal agent for primary pulmonary cryptococcosis, and we therefore recommend a six month treatment.

Infect Immun, 1999 Feb, 67(2), 936 - 41
The cell wall and membrane of Cryptococcus neoformans possess a mitogen for human T lymphocytes; Mody CH et al.; The mechanism of human T-lymphocyte activation by the pathogenic yeast Cryptococcus neoformans has not been established . Previous investigations have suggested that C . neoformans contains a mitogen for T lymphocytes, while other investigators have attributed lymphocyte proliferation in vitro to a recall antigen . Because of the potential importance of the mechanism of T-cell activation for our understanding of the immune response to C . neoformans, the present studies were performed to determine whether C . neoformans contains a mitogen for T lymphocytes . C . neoformans stimulates fetal blood lymphocytes to proliferate and stimulates proliferation of CD45RA+ cells from adults, indicating that it stimulates naive T cells . The T-cell response to C . neoformans was dependent upon the presence of accessory cells . However, allogeneic cells were sufficient for accessory cell function, indicating that the response was not major histocompatibility complex restricted . The percentage of T cells in the cell cycle was higher than that with the recall antigen tetanus toxoid but lower than that with the mitogenic lectin phytohemagglutinin A or the superantigen Staphylococcus enterotoxin B . Precursor frequency analysis established that 1 in 7,750 +/- 2, 270 T cells proliferated in response to the cryptococcal cell wall and membrane . Compared to the case for most mitogens or superantigens, the proliferative response is late and the number of T cells that enter the cell cycle and the precursor frequency are low, indicating that the mitogenic effect is modest . However, the mitogenic effect of C . neoformans should be considered when interpreting the immune response to C . neoformans, since even weak mitogens can have profound effects on host defense.

Infect Immun, 1999 Feb, 67(2), 885 - 90
Cryptococcus neoformans resides in an acidic phagolysosome of human macrophages; Levitz SM et al.; Recently, we demonstrated that human monocyte-derived macrophages (MDM) treated with chloroquine or ammonium chloride had markedly increased antifungal activity against the AIDS-related pathogen Cryptococcus neoformans . Both of these agents raise the lysosomal pH, which suggested that the increased antifungal activity was a function of alkalinizing the phagolysosome . Moreover, there was an inverse correlation between growth of C . neoformans in cell-free media and pH . These data suggested that C . neoformans was well adapted to survive within acidic compartments . To test this hypothesis, we performed studies to determine the pH of human MDM and neutrophil phagosomes containing C . neoformans . Fungi were labeled with the isothiocyanate derivatives of two pH-sensitive probes: fluorescein and 2',7'-difluorofluorescein (Oregon Green) . These probes have pKas of 6.4 and 4.7, respectively, allowing sensitive pH detection over a broad range . The phagosomal pH averaged approximately 5 after ingestion of either live or heat-killed fungi and remained relatively constant over time, which suggested that C . neoformans does not actively regulate the pH of its phagosome . The addition of 10 and 100 microM chloroquine resulted in increases in the phagosomal pH from a baseline of 5.1 up to 6.5 and 7.3, respectively . Finally, by immunofluorescence, colocalization of C . neoformans and the MDM lysosomal membrane protein LAMP-1 was demonstrated, establishing that fusion of C . neoformans-laden phagosomes with lysosomal compartments takes place . Thus, unlike many other intracellular pathogens, C . neoformans does not avoid fusion with macrophage lysosomal compartments but rather resides and survives in an acidic phagolysosome.

Nihon Kokyuki Gakkai Zasshi, 1998 Oct, 36(10), 902 - 7
{Spontaneous resolution of pulmonary cryptococcosis--report of 2 cases}; Kakeya H et al.; We encountered two patients with pulmonary cryptococcosis that resolved spontaneously without antifungal treatment . A 59-year-old man (patient 1) and 37-year-old man (patient 2) were admitted to our hospital for the examination of abnormal shadows on their chest x-ray films . Transbronchial lung biopsies were performed, and cryptococci were detected in both patient's lung tissues . Latex aggultination tests for cryptococcal antigen were positive (1:16 and 1:4, respectively) . The size of the shadows reduced spontaneously and the titers of the cryptococcal antigen in sera decreased within three months although no antifungal therapy was performed . Bronchoalveolar lavage fluid (BALF) was obtained from each patient and examined . In patient 1, the lymphocyte to CD 4/8 ratio was high in the BALF obtained on admission and the lymphocyte count was low; however, the CD 4/8 ratio was still high in a BALF sample obtained 3 months after discharge . In patient 2, the findings of the BALF analysis on admission were almost normal . The analysis of BALF may contribute to the immunological conditions to in the patients with pulmonary cryptococcosis.

J Clin Microbiol, 1999 Feb, 37(2), 315 - 20
New PCR primer pairs specific for Cryptococcus neoformans serotype A or B prepared on the basis of random amplified polymorphic DNA fingerprint pattern analyses; Aoki FH et al.; Thirty-three strains of Cryptococcus neoformans were isolated from clinical specimens, including specimens from AIDS patients in Brazil, and were classified into two serotypes; we detected 31 and 2 strains of serotypes A and B, respectively . Random amplified polymorphic DNA (RAPD) fingerprint pattern analyses of these strains of serotypes A and B showed that the patterns were similar for strains of each serotype when three 10-mer primers were used as the RAPD primers . Comparative studies of the fingerprint patterns of the study isolates with those of the reference strains also showed that the RAPD patterns for strains of each serotype were related and that most of the fingerprint bands existed commonly for all strains of each serotype tested . The common RAPD bands (an approximately 700-bp band for serotype A and an approximately 450-bp band for serotype B) were extracted and the DNA sequences were determined . Using this information, we prepared two and one PCR primer pairs which were expected to be specific for C . neoformans serotypes A and B, respectively . Use of each PCR primer combination thus prepared for serotype A or B was 100% successful in identifying the respective C . neoformans serotypes, including the 33 clinical isolates tested in the present study . Among these combinations, one for serotype A was found to amplify DNA from C . neoformans serotype B as well as serotype A . Serotype B-specific PCR primer pairs amplified DNA from not only serotype B strains but also from serotype C strains . The usefulness of other serotype-specific PCR primers for clinical C . neoformans isolates is discussed.

Diagn Microbiol Infect Dis, 1998 Nov, 32(3), 223 - 7
Evaluation of Etest for determining in vitro susceptibility of yeast isolates to amphotericin B; Pfaller MA et al.; We evaluated the performance of Etest using several different agar media for testing of amphotericin B against 660 clinical isolates of yeast species including Candida albicans, C . glabrata, C . tropicalis, C . parapsilosis, C . lusitaniae, C . krusei, Candida spp., Cryptococcus neoformans, and Saccharomyces cerevisiae . Two of the C . lusitaniae isolates represented strains with high-level amphotericin B resistance . All isolates were tested by NCCLS microdilution methods with RPMI 1640 medium and by Etest using RPMI agar with 2% glucose (RPG) . A subset of 108 isolates was also tested by Etest using RPG, antibiotic medium 3 agar (AM3), Casitone agar (CAS), and Mueller-Hinton agar (MHA) . The overall agreement between the NCCLS reference method and Etest using RPG was 98.3% . All of the Etest methods identified the two resistant strains (MICs, 4.0 to 16 micrograms/mL), whereas the reference method failed to distinguish them from 18 other isolates with MICs of 2.0 micrograms/mL . Among the 20 isolates with reference MICs of 2.0 micrograms/mL, 12 had MICs > or = 2.0 micrograms/mL when tested by Etest with RPG (range 2.0 to 16 micrograms/mL) compared with eight with AM3, two with CAS, and five with MHA . These data indicate that Etest identifies subpopulations of yeast isolates with high amphotericin B MICs . The greater sensitivity of Etest for detection of amphotericin B resistance should be exploited in future surveillance studies.

Diagn Microbiol Infect Dis, 1998 Nov, 32(3), 191 - 9
Molecular epidemiology and antifungal susceptibility of Cryptococcus neoformans isolates from Ugandan AIDS patients; Pfaller M et al.; Little is known of the antifungal susceptibility patterns and molecular epidemiology of Cryptococcus neoformans from tropical regions . We studied 164 clinical isolates of C . neofomans from 120 Ugandan AIDS patients with cryptococcal meningitis by analyzing their electrophoretic karyotypes and antifungal susceptibility profiles . Computer-assisted analysis of karyotype patterns was performed to generate dendrograms . MICs of fluconazole and flucytosine were determined by reference methods . A total of 43 distinguishable DNA types were identified among the 164 isolates . Only 30 patients (25%) were infected with their own unique strain of c . neoformans, whereas 75% of the patients shared their infecting strain with at least one other patient . Among 17 patients with more than one CSF isolate of C . neoformans, sequential isolates were identical or highly related in 12 (71%) and were different in five patients (29%) . The isolates were susceptible to both fluconazole and flucytosine and there were no instances in which a stepwise increase in either fluconazole or flucytosine MICs was observed among serial isolates . These findings suggest that the epidemiology of cryptococcal disease in AIDS patients from tropical regions may be somewhat different from that observed in more temperate climatesPIP: Even though Cryptococcus neoformans var . neoformans is a leading cause of life-threatening mycotic infection among AIDS patients worldwide, little is known about its antifungal susceptibility patterns and molecular epidemiology in tropical regions . The authors studied 164 clinical isolates of C . neoformans from 120 Ugandan AIDS patients with cryptococcal meningitis by analyzing their electrophoretic karyotypes and antifungal susceptibility profiles . Computer-assisted analysis of karyotype patterns was performed to generate dendrograms, while the MICs of fluconazole and flucytosine were determined using reference methods . 43 distinguishable C . neoformans DNA types were identified among the 164 isolates . 30 patients (25%) were infected with their own unique strain of C . neoformans, while 75% of the patients shared their infecting strain with at least 1 other patient . Among 17 patients with more than 1 cerebrospinal fluid isolate of C . neoformans, sequential isolates were identical or highly related in 12 (71%) and were different in 5 patients (29%) . The isolates were susceptible to both fluconazole and flucytosine, and there was no instance in which a stepwise increase in either fluconazole or flucytosine MICs was observed among serial isolates . These findings suggest that the epidemiology of cryptococcal disease in AIDS patients from tropical regions may be somewhat different from that observed in more temperate climates .

J Bacteriol, 1999 Jan, 181(2), 444 - 53
A glucan synthase FKS1 homolog in cryptococcus neoformans is single copy and encodes an essential function; Thompson JR et al.; Cryptococcal meningitis is a fungal infection, caused by Cryptococcus neoformans, which is prevalent in immunocompromised patient populations . Treatment failures of this disease are emerging in the clinic, usually associated with long-term treatment with existing antifungal agents . The fungal cell wall is an attractive target for drug therapy because the syntheses of cell wall glucan and chitin are processes that are absent in mammalian cells . Echinocandins comprise a class of lipopeptide compounds known to inhibit 1,3-beta-glucan synthesis, and at least two compounds belonging to this class are currently in clinical trials as therapy for life-threatening fungal infections . Studies of Saccharomyces cerevisiae and Candida albicans mutants identify the membrane-spanning subunit of glucan synthase, encoded by the FKS genes, as the molecular target of echinocandins . In vitro, the echinocandins show potent antifungal activity against Candida and Aspergillus species but are much less potent against C . neoformans . In order to examine why C . neoformans cells are less susceptible to echinocandin treatment, we have cloned a homolog of S . cerevisiae FKS1 from C . neoformans . We have developed a generalized method to evaluate the essentiality of genes in Cryptococcus and applied it to the FKS1 gene . The method relies on homologous integrative transformation with a plasmid that can integrate in two orientations, only one of which will disrupt the target gene function . The results of this analysis suggest that the C . neoformans FKS1 gene is essential for viability . The C . neoformans FKS1 sequence is closely related to the FKS1 sequences from other fungal species and appears to be single copy in C . neoformans . Furthermore, amino acid residues known to be critical for echinocandin susceptibility in Saccharomyces are conserved in the C . neoformans FKS1 sequence.

Clin Microbiol Rev, 1999 Jan, 12(1), 126 - 46
The evolutionary biology and population genetics underlying fungal strain typing; Taylor JW et al.; Strain typing of medically important fungi and fungal population genetics have been stimulated by new methods of tapping DNA variation . The aim of this contribution is to show how awareness of fungal population genetics can increase the utility of strain typing to better serve the interests of medical mycology . Knowing two basic features of fungal population biology, the mode of reproduction and genetic differentiation or isolation, can give medical mycologists information about the intraspecific groups that are worth identifying and the number and type of markers that would be needed to do so . The same evolutionary information can be just as valuable for the selection of fungi for development and testing of pharmaceuticals or vaccines . The many methods of analyzing DNA variation are evaluated in light of the need for polymorphic loci that are well characterized, simple, independent, and stable . Traditional population genetic and new phylogenetic methods for analyzing mode of reproduction, genetic differentiation, and isolation are reviewed . Strain typing and population genetic reports are examined for six medically important species: Coccidioides immitis, Histoplasma capsulatum, Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus, and A . flavus . Research opportunities in the areas of genomics, correlation of clinical variation with genetic variation, amount of recombination, and standardization of approach are suggested.

J Infect Dis, 1999 Feb, 179(2), 449 - 54
Cryptococcosis: population-based multistate active surveillance and risk factors in human immunodeficiency virus-infected persons . Cryptococcal Active Surveillance Group; Hajjeh RA et al.; To determine the incidence of cryptococcosis and its risk factors among human immunodeficiency virus (HIV)-infected persons, population-based active surveillance was conducted in four US areas (population, 12.5 million) during 1992-1994, and a case-control study was done . Of 1083 cases, 931 (86%) occurred in HIV-infected persons . The annual incidence of cryptococcosis per 1000 among persons living with AIDS ranged from 17 (San Francisco, 1994) to 66 (Atlanta, 1992) and decreased significantly in these cities during 1992-1994 . Among non-HIV-infected persons, the annual incidence of cryptococcosis ranged from 0.2 to 0.9/100,000 . Multivariate analysis of the case-control study (158 cases and 423 controls) revealed smoking and outdoor occupations to be significantly associated with an increased risk of cryptococcosis; receiving fluconazole within 3 months before enrollment was associated with a decreased risk for cryptococcosis . Further studies are needed to better describe persons with AIDS currently developing cryptococcosis in the era of highly active antiretroviral therapy.

Am J Respir Cell Mol Biol, 1999 Jan, 20(1), 135 - 42
Phenotypic and functional characterization of normal rat pleural macrophages in comparison with autologous peritoneal and alveolar macrophages; Gjomarkaj M et al.; Pleural mononuclear phagocytes (PleMP) were isolated from normal rats by pleural lavage and compared with autologous peritoneal (PerMP) and bronchoalveolar mononuclear phagocytes (BAMP) isolated by peritoneal and bronchoalveolar lavage, respectively . The phagocytic activity of PleMP, PerMP, and BAMP, evaluated by testing their ability to ingest latex beads, was lower for PleMP and PerMP than for BAMP . The phenotype of PleMP, PerMP, and BAMP was characterized by immunocytochemical staining with a panel of monoclonal antibodies (mAbs) . As expected, PleMP, PerMP, and BAMP did not react with OX19, OX33, ED5, MOM/3F12/F2, and anticytokeratin mAbs, specific for T lymphocytes, B lymphocytes, dendritic cells, granulocytes, and epithelial/mesothelial cells, respectively . Moreover, PleMP and PerMP populations were highly enriched with OX6-, OX42-, ED7-, and ED8-positive MP, whereas BAMP population was enriched with ED1- and ED9-positive cells . To test the ability of PleMP, PerMP, and BAMP to function as accessory cells (AC), mitomycin C-treated MP were used as stimulatory cells in mixed leukocyte reaction experiments, using allogeneic T cells as responders . 3HdTR incorporation by T cells was assessed as an index of AC function . PleMP and PerMP were more potent AC than BAMP . Moreover, when cultured together with autologous pulmonary interstitial dendritic cells, PleMP and PerMP exerted a more potent ability to stimulate T-cell proliferation than did BAMP . To investigate the capacity of MP to function as bactericidal and fungicidal cells, we tested their ability to kill Escherichia coli and Cryptococcus neoformans, respectively . PleMP and PerMP were less potent bactericidal and fungicidal cells than BAMP . The results of this study demonstrate that PleMP isolated from normal rat pleural space are functionally and phenotypically different from BAMP but similar to PerMP, and suggest that these cells might play an important role in cell-mediated immune reactions in the pleural space.

Arch Pathol Lab Med, 1998 Dec, 122(12), 1107 - 11
Fontana-Masson--stained tissue from culture-proven mycoses; Kimura M et al.; BACKGROUND: The Fontana-Masson stain can be used to detect melanin of dematiaceous fungi such as Curvularia lunata and Bipolaris hawaiiensis in tissue . This stain is often used to identify fungi that may not have an evident brown color in hematoxylin-eosin-stained sections . However, it is not clear whether hyphae of nondematiaceous fungi can be stained with the Fontana-Masson procedure . METHODS: Paraffin sections from culture-proven mycoses caused by species of Aspergillus, Candida, Fusarium, Pseudallescheria, Trichosporon, zygomycetes, and dematiaceous fungi were stained by the Fontana-Masson procedure . Extent of stain intensity and its distribution in fungal elements in tissue were evaluated by means of an intensity scale based on Cryptococcus neoformans as a positive control . RESULTS: Many Aspergillus spp, some zygomycetes, and all of the dematiaceous fungi showed strong intensity, whereas other fungi showed either weak intensity or no staining . CONCLUSION: Fungal hyphae other than dematiaceous fungi occasionally can be stained with the Fontana-Masson procedure . The morphology of positively stained hyphae must be carefully evaluated, especially when they are hyaline in hematoxylin-eosin-stained tissue sections.

Antimicrob Agents Chemother, 1999 Jan, 43(1), 169 - 71
In vitro activities of voriconazole, fluconazole, and itraconazole against 566 clinical isolates of Cryptococcus neoformans from the United States and Africa; Pfaller MA et al.; We investigated the in vitro activity of voriconazole compared to those of fluconazole and itraconazole against 566 clinical isolates of Cryptococcus neoformans from Africa (164) and the United States (402) . Isolates were obtained from cerebrospinal fluid (362), blood (139), and miscellaneous sites (65) . Voriconazole (MIC at which 90% of the isolates are inhibited {MIC90}, 0.12 to 0.25 microg/ml) was more active than either itraconazole (MIC90, 0.5 microg/ml) or fluconazole (MIC90, 8.0 to 16 microg/ml) against both African and U . S . isolates . Isolates inhibited by >/=16 microg of fluconazole per ml were almost all (99%) inhibited by </=1 microg of voriconazole per ml . These results suggest that voriconazole may be useful in the treatment of cryptococcosis.

Antimicrob Agents Chemother, 1999 Jan, 43(1), 106 - 14
Mixture-based heterocyclic combinatorial positional scanning libraries: discovery of bicyclic guanidines having potent antifungal activities against Candida albicans and Cryptococcus neoformans; Blondelle SE et al.; A mixture-based synthetic combinatorial library of more than 100,000 bicyclic guanidines was generated in a positional scanning format and assayed for activity against Candida albicans . Potent individual bicyclic guanidines were directly identified following the screening of the library . Time-kill curve studies indicated bactericidal activities for the individual bicyclic guanidines . These compounds also showed potent activity against Cryptococcus neoformans . These studies demonstrate the value of using mixture-based combinatorial positional scanning libraries made up of heterocyclic compounds for the rapid identification of novel classes of antifungal compounds.

Arch Pharm Res, 1998 Dec, 21(6), 688 - 91
Growth inhibitory activities of kalopanaxsaponins A and I against human pathogenic fungi; Kim DW et al.; Antifungal activities of the compounds isolated from Kalopanax pictus against representative fungi of dermatomycosis were investigated using paper disc diffusion method . It was found that kalopanaxsaponins A and I were effective in inhibiting the growth of Candida albicans KCTC 1940 and Cryptococcus neoformans KCTC 7224 with minimum inhibitory concentration (MIC) of 25 micrograms/ml . It showed that antifungal activity of both compounds have strong selectivity against the fungi of dermatomycosis.

Mayo Clin Proc, 1998 Dec, 73(12), 1205 - 25
Antifungal agents . Part I . Amphotericin B preparations and flucytosine; Patel R; Traditionally, amphotericin B has been the cornerstone of antifungal treatment . Toxicity, however, is a major dose-limiting factor of amphotericin B deoxycholate . Nevertheless, it continues to have a major role in the treatment of deep-seated mycotic infections . Recently, less nephrotic lipid formulations, including amphotericin B lipid complex, amphotericin B cholesteryl sulfate, and liposomal amphotericin B, have been introduced . The pharmacologic properties, main indications, recommended dosages, related costs, and adverse effects of these various preparations are summarized in this review . Orally administered flucytosine is useful in certain infections, particularly cryptococcal meningitis, but it should be used with caution in patients with renal insufficiency.

Infect Immun, 1999 Jan, 67(1), 220 - 9
Mechanisms for induction of L-selectin loss from T lymphocytes by a cryptococcal polysaccharide, glucuronoxylomannan; Dong ZM et al.; Disseminated cryptococcosis is accompanied by cryptococcal polysaccharides in the serum and the lack of cellular infiltrates in infected tissues . Cryptococcal polysaccharides given intravenously to mice inhibit the influx of T lymphocytes into the sites of cell-mediated immune response . The focus here was to determine whether cryptococcal polysaccharides modulate the expression of molecules, such as L-selectin, that are important in extravasation of T cells . Cryptococcal glucuronoxylomannan (GXM), but not galactoxylomannan or mannoprotein, was found to cause loss of L-selectin from freshly isolated human T cells of both CD4 and CD8 subsets and from Jurkat cells . With the signaling-pathway inhibitors staurosporine (which inhibits protein kinase C) and herbimycin A (which inhibits protein tyrosine kinases), we showed that GXM or the cryptococcal culture filtrate antigen CneF directly induces L-selectin loss from CD4(+) and CD8(+) T cells via a herbimycin A-sensitive pathway(s) presumably involving one or more protein tyrosine kinases but not via a pathway involving protein kinase C . Loss of L-selectin from the T cells before the T cells have a chance to bind to L-selectin ligands on endothelial cells would be expected to prevent T-cell migration into inflamed tissues and/or lymph organs.

Infect Immun, 1999 Jan, 67(1), 108 - 12
Catecholamine oxidative products, but not melanin, are produced by Cryptococcus neoformans during neuropathogenesis in mice; Liu L et al.; Melanin has been proposed as a virulence factor in Cryptococcus neoformans, but its presence has not been shown unambiguously in vivo . Validated methods used previously to show production of cryptococcal eumelanin pigment in vitro (P . R . Williamson, K . Wakamatsu, and S . Ito, J . Bacteriol . 180:1570-1572, 1998) were used to assess for production of laccase-derived products in mouse brain of the Lacc+ strains, 2E-TUC, H99 (serotype A), and ATCC 34873 (serotype D), and the Lacc- strain, 2E-TU . Pyrrole-2,3, 5-tricarboxylic and pyrrole-2,3-dicarboxylic acid, specific degradation products of catecholamine derivatives such as melanin, were found in all Lacc+ strains, but not in the Lacc- strain, 2E-TU . However, the presence of melanin pigment itself could not be demonstrated in the same cells . Lack of the specific degradation products aminohydroxyphenylalanine and aminohydroxyphenylethylamine in Lacc+ strains upon hydriodic acid hydrolysis showed that pheomelanin was also not produced by the fungus in vivo . These are the first data to support the generation of catecholamine oxidation products by C . neoformans in vivo, but they do not support postenzymatic polymerization of these products to form typical eumelanin, as previously proposed.

Wei Sheng Wu Xue Bao, 1997 Apr, 37(2), 135 - 41
{Polymorphism and pologenetic affinities of genotype in pathogenic yeast}; Li D et al.; The classification of pathogenic yeast, which is one of the most common reasons of opportunistic infection in human body, will be useful for the epidemiological survey and clinical works . The genotypic polymorphism of intergenus, interspecies and intraspecies in 48 isolates, including Candida, Cryptococcus, Torulopsis, Trichosporon, Saccaromyces had been administered carefully with patterns amplified by RAPD (Randomly amplified polymorphic DNA) in 53 primers . The pologenetic affinities had been evaluated by similarity coefficients obtained from these profiles . The results indicated that there are significant difference among intergenus, interspecies and intraspecies in Candida and related yeast . The similarity coefficients among Candida and Crytococcus, Trichosporon were maintained about 80% . The similarity coefficients among interspecies of Candida ranged from 82%-87%, and ones of intraspecies of different Candida species were more than 90% . The genotypic typing of species except C . guilliermondii seems to be related to morphological classification.

Mycoses, 1998 Sep-Oct, 41(7-8), 277 - 80
Fungaemia due to Cryptococcus laurentii and a review of non-neoformans cryptococcaemia; Johnson LB et al.; Cryptococcus laurentii is one of several non-neoformans cryptococci that have rarely been associated with human infection . The spectrum of clinical infection due to non-neoformans species ranges from skin lesions to fungaemia . Most cases of non-neoformans fungaemia have been nosocomially acquired and have been associated with indwelling intravascular catheters and neutropenia . Limited data on in vitro susceptibilities of non-neoformans cryptococci show these species to be more resistant to fluconazole and flucytosine than most Cr . neoformans . Two such cases are presented here.

Enferm Infecc Microbiol Clin, 1998, 16 Suppl 1, 29 - 35
{Infections by Candida and cryptococci}; Berenguer J et al.; The introduction of highly active antiretroviral therapy (HAART) has reduced dramatically the incidence of mucosal candidiasis and cryptococcosis in AIDS patients . Fluconazole is the drug of choice for candidiasis . The duration of antifungal treatment is based on response, but typically 7 to 14 days are required for oropharyngeal forms and up to 21 days for esophageal disease (200 the first day and 100 mg thereafter) . Resistant candidiasis tends to occur in persons with advanced HIV disease and previous fluconazole therapy who have been noted to result in clinical improvement with HAART . HAART must be considered the therapy of choice for refractory candidiasis . The preferred treatment for cryptococcal meningitis includes two weeks induction treatment with amphotericin B (0.7 mg/kg/d IV) with or without flucytosine (25 mg/kg qid) followed by 8 weeks of fluconazole (200-400 mg PO qd) . Long-term maintenance therapy with fluconazole (200-400 mg PO qd) is required to prevent relapses . In patients with elevated intracranial pressure who had focal neurologic deficits or mental status changes, serial lumbar punctures should be performed . In refractory cases the immediate placement of CSF drains must be considered.

Mol Cell Biol, 1999 Jan, 19(1), 745 - 50
Melanization of Cryptococcus neoformans in murine infection; Nosanchuk JD et al.; Cryptococcus neoformans is a fungus that is pathogenic in humans and that can produce melanin in vitro . Melanization is associated with virulence, but there is no evidence that melanin is made during infection . Melanins are difficult to study because they are amorphous and insoluble . Melanin-binding peptides from a phage display library were used to demonstrate that C . neoformans makes melanin-like compounds in tissue . Melanin-binding peptides were characterized by a high proportion of positively charged and aromatic residues . Two other methods, demonstration of an antibody response to melanin in mice infected with C . neoformans and analysis of yeast cell walls in infected tissue by light microscopy, were used to support these findings . The demonstration that C . neoformans melanizes in tissue has important implications for pathogenesis and drug discovery.

Anaesthesiol Reanim, 1998, 23(5), 134 - 8
{Value of high dosage fluconazole in therapy of candida infections in intensive care medicine}; Duswald KH et al.; The administration of fluconazole in the ICU setting in dosages of > or = 800 mg/day or > or = 10 mg/kg/day has been reported in about 400 patients with candidiasis of different localisation including candidemia, with a rapidly increasing incidence of serious candidal infections . In Germany, fluconazole is approved for therapy of life-threatening infections caused by Candida spp . and Cryptococcus neoformans in a dosage of up to 800 mg/day . Especially in non-neutropenic patients with life-threatening infections caused by Candida spp., Cryptococcus neoformans and Coccidioides immitis, the results of a limited number of dose-finding trials show dose-dependent response rates . These findings strongly advocate the application of high-dose fluconazole; their evaluation, however, still awaits final clarification . The good safety profile even for maximum dosages of up to 2000 mg/day and the linear, predictable pharmacokinetics up to 1600 mg/day indicate the excellent tolerability of fluconazole in the clinical situation, which justifies prospective, randomized clinical trials with treatment groups as homogeneous as possible for further evaluation of the optimum dosage and duration of treatment in the various types of candidal infection.

Ann Agric Environ Med, 1998 Jun 30, 5(1), 65 - 71
Cell wall preparations from environmental yeasts: effect on alveolar macrophage function in vitro; Sorenson WG et al.; Organic dust toxic syndrome (ODTS) is associated with inhalation of high concentrations of organic materials and is a noninfectious illness characterized by fever, malaise, myalgia, and neutrophilic inflammation of the lower respiratory tract . Studies in our laboratory of fungi in fresh lumber have demonstrated that yeasts may predominate and have raised the issue of potential exposure of sawmill workers to yeasts . Zymosan, a cell wall preparation from Saccharomyces cerevisiae, is a potent stimulator of alveolar macrophages (AM) . In the present study, preparations from the cell walls of Pichia fabianii, Candida sake, Trichosporon capitatum, Rhodotorula glutinis, and Cryptococcus laurentii were compared with zymosan and ss-1,3-glucan for their ability to stimulate AM and activate complement . All species activated complement . P . fabianii, C . sake, T . capitatum, R . glutinis, C . laurentii, as well as zymosan and glucan, stimulated superoxide anion and leukotriene B4 production in a dose-dependent fashion, but R . glutinis and C . laurentii were much less active . Zymosan, glucan, P . fabianii, and R . glutinis treatment of AM resulted in increased phagocytosis of labeled sheep RBCs, whereas there was no effect with C . sake or C . laurentii and T . capitatum significantly inhibiting phagocytosis . These results suggest that exposure to high concentrations of yeast could provoke pulmonary inflammation resulting in an episode of ODTS.

Mycopathologia, 1998, 142(1), 3 - 7
Anticryptococcal activity of voriconazole against Cryptococcus neoformans var . gatti vs var . neoformans: comparison with fluconazole and effect of human serum; Brummer E et al.; Voriconazole (VCZ), a new wide-spectrum antifungal triazole currently in development, was tested for activity against Cryptococcus neoformans (CN) var . gattii and var . neoformans in RPMI-1640 (RPMI) or RPMI plus human serum . In RPMI VCZ was 10-fold more inhibitory than FCZ for both varieties of CN . In the presence of human serum neither VCZ nor FCZ had enhanced activity against CN var . gattii . By contrast, both VCZ and FCZ had significantly increased activity in the presence of serum against CN var . neoformans . The lack of serum-enhancing activity for VCZ or FCZ against CN var . gattii may reflect the in vivo situation and predict less efficacy in CN var . gattii infections.

Nihon Kokyuki Gakkai Zasshi, 1998 Aug, 36(8), 708 - 12
{Primary pulmonary cryptococcosis with pleural effusion}; Matsunaga K et al.; A 32-year woman presented at our hospital with left chest pain . Chest X-ray films revealed multiple nodular shadows (accompanied by a fusing tendency) and pleural effusion in the left lower lung field . Transbronchial lung biopsy specimens contained cryptococcal fungi . Blood and pleural effusion samples were positive for cryptococcal antigen, but cerebrospinal fluid was not . Because cryptococcus was not detected in cultures of cerebrospinal fluid, urine, or blood, and because the patient had no underlying disease, she was given a diagnosis of primary pulmonary cryptococcosis . On the second consultation 10 days later, the patient's subjective symptoms had subsided and the X-ray findings were generally improved . The condition resolved spontaneously without treatment within 4.5 months . This case is noteworthy for several reasons: (1) pleural effusion was observed, and the fluid could be examined: (2) the inflammatory reactions were pronounced at the time of first examination; and (3) the disease developed abruptly but subsided spontaneously within a short period of time . We reviewed 15 cases of primary pulmonary cryptococcosis reported recently in this journal, including this case . That review suggests that the finding concerning inflammatory reactions are associated with the motive of detection during initial examination.

Mt Sinai J Med, 1998 Oct-Nov, 65(5-6), 422 - 5
Value of assessing cryptococcal antigen in bronchoalveolar lavage and sputum specimens from patients with AIDS; Bottone EJ et al.; Cryptococcus neoformans has become a significant opportunistic pathogen, accounting for 8-10% of infectious complications in patients with AIDS . When encapsulated yeast cells are observed in Giemsa-stained smears of bronchoalveolar washings (BAL), or induced sputum specimens, confirmation as C . neoformans is germane to definitive therapy . We therefore studied 30 BAL and 9 induced sputum specimens for cryptococcal antigen . Of the 30 BAL, 3 specimens were positive for cryptococcal antigen, ranging in titer from 1:4 to 1:256, and 2 of 9 sputum samples were also smear, culture and antigen positive (titer 1:2) for C . neoformans . Of the 34 negative specimens, none of the seven containing Candida species or the one containing H . capsulatum or the one containing P . carinii cross-reacted with cryptococcal anticapsular antibody . Our results indicate that when yeast forms suggestive of C . neoformans are visualized on direct smears of BAL or sputum samples, rapid confirmation as C . neoformans may be achieved by assessment for cryptococcal antigen . A correlation may also exist between antigen titer in respiratory specimens and extent of cryptococcal infection.

An Med Interna, 1998 Oct, 15(10), 523 - 7
{Fungemia in patients with HIV infection}; Santos J et al.; OBJECTIVE: To make an analysis of fungemia in HIV-infected patients in our hospital . PATIENTS AND METHODS: We retrospectively (1989-1997) studied all HIV-infected patients with positive blood cultures for Candida sp., Cryptococcus neoformans or any other fungal infection . RESULTS: C . neoformans was isolated in 11 patients (10 men and 1 woman): Six were treated with amphotericin B and 5 with fluconazole . 2 patients died during the acute phase and the infection relapsed in 3 . Blood culture for Candida sp . were positive in 9 (8 men and 1 woman): only a case was nosocomial . Seven patients were intravenous drug users and the presenting manifestations were autolimited candidemia in 3, aortic and tricuspid endocarditis in 1 and 2 cases respectively and pneumonia in another one . Six C . albicans, 2 C . krusei and 1 C . glabrata were isolated . 3 patients received amphotericin B and 3 received fluconazole . 2 patients suffering from endocarditis died and so did the patient with C . glabrata infection . A patient, who denied having travelled to endemic areas, developed histoplasmosis; blood culture was positive for H . capsulatum . He initially had a good response to amphotericin B and itraconazole . CONCLUSIONS: Fungemia is not frequent in HIV-infected patients . Cryptococcosis and histoplasmosis occur in advanced HIV-patients and candidemia is fundamentally associated with intravenous drug use.

Proc Natl Acad Sci U S A, 1998 Dec 8, 95(25), 14967 - 72
Phenotypic switching in the human pathogenic fungus Cryptococcus neoformans is associated with changes in virulence and pulmonary inflammatory response in rodents; Goldman DL et al.; High-frequency reversible changes in colony morphology were observed in three strains of Cryptococcus neoformans . For one strain (SB4, serotype A), this process produced three colony types: smooth (S), wrinkled (W), and serrated (C) . The frequency of switching between colony types varied for the individual colony transitions and was as high as 10(-3) . Mice infected with colony type W died faster than those infected with other colony types . The rat inflammatory response to infection with colony types S, W, and C was C > S > W and ranged from intense granulomatous inflammation with caseous necrosis for infection with type C to minimal inflammation for infection with type W . Infection with the various colony types was associated with different antibody responses to cryptococcal proteins in rats . Analysis of cellular characteristics revealed differences between the three colony types . High-frequency changes in colony morphology were also observed in two additional strains of C . neoformans . For one strain (24067A, serotype D) the switching occurred between smooth and wrinkled colonies . For the other strain (J32A, serotype A), the switching occurred between mucoid and nonmucoid colonies . The findings indicate that C . neoformans undergoes phenotypic switching and that this process can affect virulence and host inflammatory and immune responses . Phenotypic switching may play a role in the ability of this fungus to escape host defenses and establish chronic infections.

Rev Cubana Med Trop, 1998, 50(1), 27 - 30
{Isolation of anti-Cryptococcus neoformans immunoglobulin G}; Illnait Zaragozi MT et al.; Two immunization schemes were applied to rabbits with an autoctonous strain aimed at obtaining an anti-Crytococcus neoformans IgG . Whole cells treated with formalin were used . Dosages and immunization routes were different . The sera obtained were titered and that with the highest titer was selected (1:1 024 by the laminar agglutination method) and purified by saline precipitation and ion exchange chromatography . An IgG of high purity and biological activity was obtained.

Folia Neuropathol, 1998, 36(3), 129 - 44
Opportunistic infections of the central nervous system in the course of acquired immune deficiency syndrome (AIDS) . Morphological analysis of 172 cases; Zelman IB et al.; A neuropathological analysis of 172 cases of AIDS in adults was carried out, to determine the occurrence and nature of the opportunistic infections of the central nervous system (CNS) . The material comprised 155 cases of men, and 17 women . Mean age of patients was 38 years . Collection under study originated from the period between 1987 and 1997 . Opportunistic infections were present in 57.5 percent of cases being in 38.4 percent the only pathological process, whereas in 19.1 percent they coexisted with HIV-dependent pathology or with neoplastic growth . Cytomegalovirus infection (22.7%), toxoplasmosis (16.3%), cryptococcosis (8.1%) and progressive multifocal leukoencephalopathy (9.3%) were the most common opportunistic infections of CNS . The remaining viral (herpetic encephalitis, tick-borne encephalitis and herpes zoster multifocal encephalitis), bacterial (lues, metastatic encephalitis connected with heart valvular changes) and fungal (candidiasis) infections were present only in single cases . It is worth mentioning 3 cases of brain aspergillosis and 5 cases of leptomeningeal tuberculosis . Great morphological variability in the most common opportunistic infections found in our material (cytomegaly, toxoplasmosis, cryptococcosis and PML) was the most striking phenomenon . Neuropathological abnormalities in cases of toxoplasmosis and cryptococcosis revealed remarkable dependence on clinical medication used . Cases of PML were characterized by strong variances of the type and intensity of demyelination, ranging from disseminated foci of various size to diffuse complete myelin loss in the white matter involving uni- or bilaterally cerebral or cerebellar hemispheres . The coexistence of opportunistic infections with HIV-dependent cerebral pathology or other types of opportunistic processes was a very characteristic feature . Concomitance of HIV-dependent pathology with viral opportunistic processes was common . The frequency of this concomitance and more severe HIV-dependent pathology in cases with other viral cerebral infections may suggest pathogenetic interaction of viral infections . Cerebral tuberculosis was less frequent as compared with other neuropathological collections, especially those from the United States . However, it seems worth mentioning that 3 of 5 cases occurred in the last year of observation.

Eur J Clin Microbiol Infect Dis, 1998 Sep, 17(9), 637 - 41
Comparison of five days versus seven days of incubation for detection of positive blood cultures by the Bactec 9240 system; Huang AH et al.; The specimen capacity of blood culture systems is determined by the length of time that blood cultures are incubated . Since the patient populations served by hospitals vary, individual laboratories should evaluate the relative cost and benefits of different testing regimens for their particular setting . To be cost-effective, the use of a 5-day rather than a 7-day protocol for the Bactec 9240 system (Becton Dickinson Diagnostic Instrument Systems, USA) has been recommended . To evaluate whether the shorter schedule would be appropriate at the Microbiology Laboratory at the National Cheng Kung University Medical Center in Tainan, Taiwan, the yield from 5 days versus 7 days of incubation was compared using a total of 9653 blood specimens collected from 1 April to 30 September 1997 . Of the 1848 positive vials, 1822 (98.6%) were positive in the first 5 testing days; only 26 (1.4%) were positive on day 6 or 7 . Only five of the latter vials were judged to contain clinically significant organisms: one Cryptococcus neoformans, one Candida albicans, one Enterobacter cloacae, one Klebsiella pneumoniae, and one Proteus mirabilis . Two vials contained organisms whose clinical significance was categorized as unknown, while 19 contained organisms that were considered contaminants . These data suggest that, for the laboratory studied, the 5-day protocol for the Bactec 9240 system is more cost-effective than the 7-day protocol and is adequate for detection of positive blood cultures.

Rev Inst Med Trop Sao Paulo, 1998 May-Jun, 40(3), 125 - 35
Virulence factors in fungi of systemic mycoses; Kurokawa CS et al.; Pathogenic fungi that cause systemic mycoses retain several factors which allow their growth in adverse conditions provided by the host, leading to the establishment of the parasitic relationship and contributing to disease development . These factors are known as virulence factors which favor the infection process and the pathogenesis of the mycoses . The present study evaluates the virulence factors of pathogenic fungi such as Blastomyces dermatitidis, Coccidioides immitis, Cryptococcus neoformans, Histoplasma capsulatum and Paracoccidioides brasiliensis in terms of thermotolerance, dimorphism, capsule or cell wall components as well as enzyme production . Virulence factors favor fungal adhesion, colonization, dissemination and the ability to survive in hostile environments and elude the immune response mechanisms of the host . Both the virulence factors presented by different fungi and the defense mechanisms provided by the host require action and interaction of complex processes whose knowledge allows a better understanding of the pathogenesis of systemic mycoses.

J Formos Med Assoc, 1998 Oct, 97(10), 690 - 7
Bacteremia and fungemia in patients with advanced human immunodeficiency virus (HIV) infection in Taiwan; Hung CC et al.; To understand the etiology and clinical outcome of bacterial and fungal sepsis in patients with advanced human immunodeficiency virus (HIV) infection in Taiwan, we conducted a prospective study of nonmycobacterial bacteremia and fungemia in HIV-infected patients with fever who were admitted to a university hospital in Taiwan during a 42-month period . Of 210 patients, 41 (19.5%) had a total of 52 episodes of sepsis due to nonmycobacterial bacteria or fungi, or both (15.5% of 336 episodes of fever) . All but one patient had acquired immunodeficiency syndrome (AIDS), and the mean CD4 lymphocyte count was 29/microL (range, 0-321/microL) . A total of 57 pathogens (39 bacteria and 18 fungi) were isolated from blood; polymicrobial sepsis due to both bacteria and fungi occurred in four episodes . Nontyphoid Salmonella (NTS) was the most common cause of community-acquired bacteremia (24/30, 80%) . Staphylococcus aureus bacteremia was diagnosed in three episodes while Streptococcus pneumoniae bacteremia was found in only one . Cryptococcus neoformans was the most common cause of fungemia and was responsible for 12 episodes, while fungemia due to Penicillium marneffei and Histoplasma capsulatum, two emerging fungi in Taiwan, were diagnosed in four cases and one case, respectively . Nine episodes, eight of bacteremia and one of candidemia, were nosocomial . The overall in-hospital mortality was 29%, and nosocomial sepsis was associated with a higher mortality rate (56%, p = 0.02) . The mean duration of survival after recovery from initial sepsis was 426 days . We conclude that NTS bacteremia was the most common cause of sepsis in patients with advanced HIV infection in Taiwan and clinicians caring for such patients should watch for emerging fungal infections . Nosocomial sepsis was associated with a high mortality rate . The mean survival duration after recovery from sepsis of our patients was short.

Clin Infect Dis, 1998 Nov, 27(5), 1231 - 7
Acute respiratory failure associated with cryptococcosis in patients with AIDS: analysis of predictive factors; Visnegarwala F et al.; The incidence of acute respiratory failure (ARF) associated with cryptococcal disease in patients with AIDS is underestimated in the literature . We performed a retrospective, case-control (referent) study to determine the prevalence of ARF associated with cryptococcal disease and analyzed associated factors . Potential cases of ARF were identified at four university-affiliated teaching hospitals from a cohort of 210 patients with AIDS who had positive cryptococcal antigen tests and/or Cryptococcus neoformans isolated from any body site . Twenty-nine of the 210 (13.8%) had ARF associated with cryptococcal disease . Nineteen were thought to have respiratory failure due solely to C . neoformans . The demographic, clinical, laboratory, treatment, and outcome data of 19 cases of respiratory failure were compared with data for 20 patients without respiratory failure . In-hospital mortality was 100% and median survival was 2 days for cases, vs . 25% and > 365 days, respectively, for referents . The clinical presentation was identical to that of Pneumocystis carinii pneumonia . In multivariate analysis, variables independently predictive of ARF in patients with cryptococcal disease were black race, a lactate dehydrogenase level of > or = 500 IU/L, the presence of interstitial infiltrates, and the presence of cutaneous lesions . ARF with cryptococcosis in patients with AIDS is associated with disseminated disease and high mortality . The diagnosis frequently is not considered before death . Serum cryptococcal antigen testing is a sensitive and rapid screening method.

Clin Infect Dis, 1998 Nov, 27(5), 1138 - 47
The epidemiological features of invasive mycotic infections in the San Francisco Bay area, 1992-1993: results of population-based laboratory active surveillance; Rees JR et al.; Population-based active laboratory surveillance for invasive mycotic infections was conducted during 1992 and 1993 in three California counties: Alameda, Contra Costa, and San Francisco (population, 2.94 million) . The cumulative incidence of invasive mycotic infections was 178.3 per million per year . Invasive mycoses were most commonly caused by Candida (72.8 per million per year), Cryptococcus (65.5), Coccidioides (15.3), Aspergillus (12.4), and Histoplasma (7.1) . The clinical significance of other, less common fungi was determined by detailed chart review . The cumulative incidence was determined for zygomycosis (1.7 per million per year), hyalohyphomycosis (1.2), and phaeohyphomycosis (1.0) . The most common underlying conditions were human immunodeficiency virus infection (47.4%), nonhematologic malignancy (14.7%), diabetes mellitus (9.9%), and chronic lung disease (9.3%) . This represents the first population-based epidemiological assessment of invasive mycoses in the United States.

J Am Anim Hosp Assoc, 1998 Nov-Dec, 34(6), 523 - 6
Cervical spinal cord compression caused by cryptococcosis in a dog: successful treatment with surgery and fluconazole; Kerwin SC et al.; A six-year-old, male Doberman pinscher was presented for acute onset of upper motor neuron tetraparesis . An extradural compressive lesion compatible with intervertebral disk rupture at the sixth to seventh cervical (C6-C7) disk space was evident on myelography . A large, gelatinous mass of pure cryptococcal organisms causing spinal cord compression was identified upon exploratory surgery . Removal of the mass caused relief of clinical signs . No evidence of involvement of other organ systems was found; however, serum and cerebrospinal fluid titers were positive for cryptococcal infection . The dog was treated with fluconazole (5.5 mg/kg body weight, per os sid) until serum titers for cryptococcal infection were negative at seven months postsurgery . To the authors' knowledge, this is the only report of a dog with cryptococcosis treated successfully using fluconazole as a sole agent.

Crit Rev Oral Biol Med, 1998, 9(4), 480 - 97
Human salivary histatins: promising anti-fungal therapeutic agents; Tsai H et al.; Histatins constitute a group of small, cationic multifunctional proteins present in the saliva of human and some non-human primates . The most significant function of histatins may be their anti-fungal activity against Candida albicans and Cryptococcus neoformans . Histatins have been extensively studied at both the protein and gene levels . The structure-function relationship of histatins with respect to their candidacidal activity has also been studied by means of recombinant histatin variants, as well as by chemically synthesized histatin fragments . The mechanism of histatins' action on Candida albicans is not clear, but it appears to be different from that of azole-based anti-fungal drugs which interrupt ergosterol synthesis . During the past 20 years, fungal infections have become more prevalent as a result of the emergence of AIDS, as well as, paradoxically, modern medical advances . The toxicity of current anti-fungal medicine, the emergence of drug-resistant strains, and the availability of only a few types of anti-fungal agents are the major disadvantages of current anti-fungal therapy . Therefore, the importance of the search for new, broad-spectrum anti-fungals with little or no toxicity cannot be overemphasized . The following properties make histatins promising anti-fungal therapeutic agents: (1) They have little or no toxicity; (2) they possess high cidal activities against azole-resistant fungal species and most of the fungal species tested; and (3) their candidacidal activity is similar to that of azole-based antifungals . Current research efforts focus on the development of improved histatins with enhanced cidal activity and stability, and of suitable and effective histatin delivery systems . These and other approaches may help to outpace the growing list of drug-resistant and opportunistic fungi causing life-threatening, disseminating diseases . The histatins with improved protective properties may also be used as components of artificial saliva for patients with salivary dysfunction.

J Infect, 1998 Sep, 37(2), 112 - 5
A prospective study of Glasgow Coma Scale (GCS), age, CSF-neutrophil count, and CSF-protein and glucose levels as prognostic indicators in 100 adult patients with meningitis; Schutte CM et al.; BACKGROUND: The Glasgow coma scale (GCS) is an objective measurement of a patient's level of consciousness and has prognostic implications in traumatic head injuries . Morbidity and mortality of patients with meningitis have been related amongst others to level of consciousness, hypoglycorrhachia, extremes of age, and high CSF protein values . In this prospective study of 100 patients the correlation between the GCS, age, CSF-neutrophil count and CSF-glucose and protein levels and the eventual outcome of the patients was assessed . METHODS: In 100 consecutive patients with meningitis (bacterial, viral, tuberculous, cryptococcal and other) the GCS, age, CSF-neutrophil count and CSF-protein and glucose levels were determined at admission . After treatment the outcome of the patient was assigned to one of four categories: healthy, minor and severe neurological deficits and death . RESULTS: From a non-parametric one-way analysis of variance it was found that with respect to mean GCS-values significant differences were present among the outcome categories (P < 0.0001) . The outcome categories did not differ significantly with respect to age, CSF-neutrophil count or CSF-glucose level, but did differ significantly with respect to the CSF-protein level (P < 0.0025) . Additionally, 88% of patients with a GCS value of > 12 had a good neurological outcome, while 88% of those with a GCS value of < or = 8 had a poor outcome . CONCLUSION: A good correlation between both the GCS and CSF-protein level at admission and the outcome of patients with meningitis was found, with the GCS value being a better prognostic indicator than high CSF protein levels.

Chest, 1998 Nov, 114(5), 1258 - 63
The effect of adjunctive corticosteroids for the treatment of Pneumocystis carinii pneumonia on mortality and subsequent complications; Gallant JE et al.; OBJECTIVE: To assess the long-term safety of adjunctive corticosteroids in the treatment of Pneumocystis carinii pneumonia (PCP) . DESIGN: Analysis of data from a large prospective observational database . SETTING: HIV clinic at a large urban teaching hospital . PATIENTS: One hundred seventy-four patients who developed PCP after being enrolled in the database . RESULTS: Fifty-three patients (30%) received adjunctive corticosteroids and 121 (70%) did not . Survival did not differ between groups after adjusting for CD4 count (relative risk for adjunctive corticosteroids = 0.74, p = 0.13) . There were no differences in the incidence of cytomegalovirus disease (adjunctive corticosteroids: 18.5 cases per 100 person-years vs no adjunctive corticosteroids: 15.7, p = 0.22), Mycobacterium avium complex (23.4 vs 27.0, p = 0.73), cryptococcal meningitis (1.8 vs 4.1, p = 0.58), toxoplasmosis (3.6 vs 11.0, p = 0.28), Kaposi's sarcoma (1.8 vs 2.2, p = 0.92), herpes simplex (27.1 vs 42.7, p = 0.66), herpes zoster (3.8 vs 6.9, p = 0.71), oropharyngeal candidiasis (18.9 vs 10.9, p = 0.09), or non-Hodgkin's lymphoma (3.5 vs 4.2, p = 0.92) . Esophageal candidiasis was more common among adjunctive corticosteroid recipients (45.1 vs 26.6, p = 0.01) . Results were similar for time to development of opportunistic conditions . CONCLUSIONS: Adjunctive corticosteroids do not increase mortality or the risk of most common HIV-associated complications.

J Antibiot (Tokyo), 1998 Sep, 51(9), 829 - 36
Fermentation studies of rustmicin production by a Micromonospora sp; Sigmund JM et al.; The antifungal antibiotic rustmicin was detected in the fermentation broth of the actinomycete MA 7094 as a specific inhibitor of sphingolipid biosynthesis in Candida albicans and as a potent fungicidal agent against Cryptococcus neoformans . Taxonomic characterization by both classical means and PCR fingerprinting supported the assignment of the producing culture to the genus Micromonospora . Fermentation medium optimization studies showed that the concentration of tomato paste in the medium was critical to increased production of rustmicin by MA 7094 . The stimulatory effect of tomato paste in the medium on rustmicin production appeared to be related to the maintenance of pH at or below a value of 6.0 . Addition of the antifoam agent P-2000 to the fermentation was found to dramatically reduce the rustmicin titer, while substitution of another antifoam agent, UCON-LB625, resulted in a 100% increase in the amount of rustmicin detected . After fermentation optimization studies and the generation of a non-sporulating mutant of MA 7094, the rustmicin titer was increased from an initial titer of 10mg/liter to 145 mg/liter.

J Infect, 1998 Jul, 37(1), 36 - 8
A retrospective study on the efficacy and safety of amphotericin B in a lipid emulsion for the treatment of cryptococcal meningitis in AIDS patients; Torre D et al.; To evaluate the efficacy and safety of Amphotericin B dissolved in dextrose (Amb) or in a lipid emulsion (Intralipid, Amb-IL) in AIDS patients with cryptococcal meningitis, we conducted a retrospective study in 30 AIDS patients with cryptococcal meningitis . A clinical complete resolution was obtained in 11 patients (55%) treated with Amb, and in six patients (60%) treated with Amb-IL . Intralipid did not decrease the infusion-related adverse effects, in particular nephrotoxicity and anaemia . Our results indicate that Amb-IL formulation is useful in the treatment of cryptococcal meningitis in AIDS patients, but it does not reduce the infusion-related adverse events.

J Infect, 1998 May, 36(3), 329 - 31
Microscopy and latex antigen negative cryptococcal meningitis; Laurenson IF et al.; A HIV-positive patient presented with cryptococcal meningitis that was not detected by cerebrospinal fluid (CSF) latex antigen and direct microscopy . The diagnosis was confirmed by culture of the CSF and subsequent urine culture, both of which yielded an apparently acapsular strain of Cryptococcus neoformans . After 19 months the patient relapsed and capsulated yeasts were observed on this occasion on direct microscopy of the CSF . The latex antigen test was strongly positive . Culture again yielded an apparently acapsular isolate . Retrospective culture of all isolates obtained from this patient in sterile CSF resulted in the formation of capsules . This was confirmed by the requirement of normal non heat inactivated serum for neutrophil-cryptococcus attachment to occur in vitro . Although antigen and direct microscopy are frequently relied upon to diagnose cryptococcal meningitis, a negative result does not exclude the condition.

J Infect, 1998 Mar, 36(2), 231 - 2
Meningitis due to a combined infection with Cryptococcus neoformans and Streptococcus pneumoniae in an AIDS patient; John MA et al.; We report on a case of dual infection with Cryptococcus neoformans and Streptococcus pneumoniae in the cerebrospinal fluid of a patient with AIDS . To our knowledge this represents the first case to be reported in the English literature . In immunocompromised patients the importance of performing both the Gram- and India-ink stain and cryptococcal antigen test on CSF specimens with completely normal biochemical and cellular parameters is emphasizedPIP: This report presents the first case described in the English literature of dual infection with Cryptococcus neoformans and Streptococcus pneumoniae in the cerebrospinal fluid of an AIDS patient . The patient was a 32-year-old, HIV-positive South African woman who had been diagnosed with disseminated tuberculosis 5 months prior to the index admission . Her chief complaints at presentation were abdominal pain, chronic diarrhea, and vaginal discharge, suggesting a diagnosis of pneumonia and pelvic inflammatory disease . Persistence of confusion led to a lumbar puncture; gram-positive and budding yeasts were observed and subsequent India-ink staining revealed capsulated yeast typical of C . neoformans . S . pneumoniae and C . neoformans were cultured 24 and 48 hours, respectively, after incubation . The woman died within 24 hours of hospital admission, precluding further investigation . It is presumed that this woman already had disseminated cryptococci at the earlier presentation, but was incorrectly diagnosed as having tuberculosis . The finding of pneumococci, in the absence of inflammatory cells in cerebrospinal fluid, suggests the terminal event was fulminant pneumococcal meningitis in the setting of chronic cryptococcal meningitis . This case supports the importance of performing both the Gram- and India-ink stains and cryptococcal antigen test on cerebrospinal fluid specimens from immunocompromised patients, even when biochemical and cellular parameters are normal .

J Infect, 1998 Mar, 36(2), 220 - 2
Cryptococcosis: an unusual opportunistic infection complicating B cell lymphoproliferative disorders; Melzer M et al.; We report two cases of cryptococcosis in patients with Waldenstrom's macroglobulinaemia and chronic lymphocytic leukaemia that responded to prolonged therapy with systemic amphotericin and flucytosine . Cryptococcosis, although more common in those with impaired cell mediated immunity, should also be considered as a complication in patients with impaired antibody responses.

J Infect, 1998 Jan, 36(1), 118 - 9
A child case of haemophagocytic syndrome associated with cryptococcal meningoencephalitis; Numata K et al.; A previously healthy 12-year-old Japanese girl developed meningoencephalitis due to Cryptococcus neoformans . During the course of her illness she suffered persistent high fever, severe pancytopenia, hypercytokinemia and liver dysfunction . Laboratory findings, including results of a bone marrow examination, strongly indicated complication by haemophagocytic syndrome (HPS) . The preceding cryptococcal infection was thought to be a cause of the HPS because no other viral or bacterial infection could be confirmed . The girl died of acute respiratory failure during the progressive course of HPS . This may be the first reported case of HPS due to cryptococcal infection in an otherwise healthy child.

J Clin Microbiol, 1998 Dec, 36(12), 3653 - 6
Variation in electrophoretic karyotype and antifungal susceptibility of clinical isolates of Cryptococcus neoformans at a university-affiliated teaching hospital from 1987 to 1994; Klepser ME et al.; Ninety-eight isolates of Cryptococcus neoformans were collected from 30 patients at the University of Iowa Hospitals and Clinics from December 1987 through December 1994 . The susceptibility of each isolate was determined against fluconazole, itraconazole, amphotericin B, and flucytosine . Of the 98 isolates, 53 were recovered from blood, 19 were recovered from cerebrospinal fluid (CSF), and 26 were recovered from other sources . Although the strains were isolated from the same institution, DNA typing by electrophoretic karyotype (EK) revealed wide genetic variation . Overall, 23 different EK profiles were identified by computer-aided analysis . An isolate exhibiting a single EK was isolated from 24 of 30 patients (80%), whereas multiple strains with unique EKs were isolated from 6 of 30 (20%) patients . Of the six patients who had multiple strains recovered, only one individual had two strains isolated from unique body sites, one strain from the blood and the other from the CSF . Six strains were isolated from multiple patients . Nine patients had multiple sequential isolates recovered over periods of time ranging from 3 days to 4 months . EK analysis revealed persistence of the same genotype in six of the cases . Three patients, however, appeared to have an isolate with a second distinct EK emerge during therapy . Of the patients with sequential positive cultures, an increase in the MICs for test agents was observed in only one case . C . neoformans isolates were collected over a period of 7 years, during which time MICs at our institution remained stable.

J Infect Dis, 1998 Dec, 178(6), 1761 - 6
Serial isolates of Cryptococcus neoformans from patients with AIDS differ in virulence for mice; Fries BC et al.; Serial isolates of Cryptococcus neoformans from patients with chronic infection can exhibit minor karyotype changes as a result of chromosome length polymorphism (CLP) . This study investigated whether serial C . neoformans isolates with CLP from 4 patients with AIDS exhibited biologic and phenotypic differences . CLP permits the identification of serial isolates in murine mixed infection . The parameters studied were virulence in mice, capsule size, colony morphology, melanization, protease production, MICs of antifungal drugs, and growth rates in vitro . Two parameters of virulence in mice were studied: persistence in tissue and survival time after lethal infection . Serial C . neoformans isolates were shown to differ in ability to persist in vivo, virulence in a murine infection model, in vitro growth rates at 37 degreesC, and capsule size . Melanin and protease production and MICs of antifungal drugs were comparable for serial isolates . These observations suggest microevolution of C . neoformans during human infection . This process may allow the fungal population to change, escape eradication by the immune system, and thus cause chronic infections.

Zentralbl Bakteriol, 1995 Nov, 283(1), 127 - 35
CD4 lymphocyte count in HIV-positive persons exposed to Cryptococcus neoformans; Arasteh K et al.; A report is presented on four HIV-positive homosexual men examined after several months of exposure during cleaning of a flat from masses of pigeon droppings heavily colonized by Cryptococcus neoformans . Only one out of the four persons, with a CD4 lymphocyte count of 50/microL, fell sick from systemic cryptococcosis, but not the others, with CD4 lymphocyte counts of 180, 250, and 630/microL, respectively; they remained clinically and mycologically inconspicuous and free from C . neoformans . Open questions in view of the epidemiology of opportunistic pathogens in AIDS are discussed with regard to the CD4 cell count as a parameter indicating a predisposition for cryptococcosis as an airborne AIDS-defining opportunistic infection . This has been confirmed by specific cultural diagnosis of the agent in both the environment and the patient . Already in 1987/88, the probable source of infection had been the subject of epidemiological studies on C . neoformans in Berlin.

Fungal Genet Biol, 1998 Oct, 25(1), 1 - 14
Signal transduction pathways regulating differentiation and pathogenicity of Cryptococcus neoformans; Alspaugh JA et al.; The basidiomycetous yeast Cryptococcus neoformans is a human pathogen . Several phenotypes of this organism are defined as virulence traits including the polysaccharide capsule, melanin, and the ability to grow at 37 degreesC . The signaling pathways regulating the expression of these phenotypes and other important cellular processes are being defined on a molecular level . For example, the highly conserved signaling molecule calcineurin regulates high temperature growth in C . neoformans . A cryptococcal homolog of Saccharomyces cerevisiae STE12, the gene for a transcriptional regulator activated by the MAP kinase cascade, has also been identified . Additionally, the C . neoformans Galpha protein GPA1 and cAMP regulate mating, melanin production, encapsulation, and pathogenicity . This fungus is an excellent model to further dissect virulence-associated signaling pathways . The conserved role of Galpha proteins and cAMP-associated signaling pathways in fungal differentiation and pathogenicity is also reviewed .

J Infect Dis, 1998 Oct, 178(4), 1213 - 6
A human IgM monoclonal antibody prolongs survival of mice with lethal cryptococcosis; Fleuridor R et al.; Antifungal therapy cannot eradicate Cryptococcus neoformans infections in immunosuppressed patient groups . Therefore, adjunctive antibody-based therapy is being considered to enhance host immune responses to C . neoformans . To characterize potentially protective reagents, the idiotypic repertoire of human antibodies to cryptococcal glucuronoxylomannan (GXM) elicited by the investigational conjugate vaccine GXM-tetanus toxoid was examined . The variable genes used by human antibodies to GXM were analyzed with an antigen-based ELISA and mouse monoclonal antibodies (MAbs) that recognize determinants of human VH1, VH3, and VH4 gene segments . Antibodies to GXM were shown to use VH3 gene segments, and antibodies with the greatest binding to GXM also bound to protein A . A VH3-positive human monoclonal IgM prolonged survival of C . neoformans-infected mice . This is the first report that a human antibody is protective against C . neoformans . These results suggest that human MAbs may have promise as therapeutic reagents against cryptococcosis.

Rev Chir Orthop Reparatrice Appar Mot, 1998 Sep, 84(5), 451 - 5
{Osseous site of systemic cryptococcosis in an immunocompetent patient}; Goncalves J et al.; Cryptococcosis is a cosmopolite mycosis caused by an opportunistic fungus usually infecting immunodeficient subjects . A 50-year old patient, with no particular medical history was seen with metaphyseal and diaphyseal bony lesions of a non-specific lytic aspect . This demonstrates that such an infection may also occur in apparently immunocompetent subjects . Diagnosis may be difficult and needs a special investigation looking for Cryptococcus neoformans . It should perhaps be made when first-intention investigations of a lytic bony lesion does not reveal any precise aetiology.

J Med Assoc Thai, 1998 Nov, 81(11), 893 - 905
Prolonged fever due to Mycobacterium avium complex (MAC) disease in advanced HIV infection: a public health concern; Suwanagool S et al.; From March 1997 to June 1998, infectious etiologies of prolonged fever was prospectively investigated in 104 advanced human immunodeficiency virus (HIV) infected patients admitted to Siriraj Hospital . The etiology could be identified in 91 cases (87.5%) . Of these, blood cultures from 68 patients yielded mycobacteria and fungi . Mycobacterium avium complex was the most common blood isolate in 24 per cent of the patients; followed by Mycobacterium tuberculosis in 20.2 per cent, Cryptococcus neoformans in 5.8 per cent, Penicillium marneffei in 5.8 per cent . During the course of febrile illness, 79 of the 91 patients (86.8%) exhibited focal lesions . Weight loss, elevated serum alkaline phosphatase were often found to be significantly more associated with MAC bacteremia (P < 0.05) . Pulmonary involvement significantly correlated more with M . tuberculosis bacteremia than MAC bacteremia (P < 0.05) . No cause could be identified in 13 cases . Mycobacterium blood culture alone established the etiologies in 68 cases (65.4%) . Of the 25 patients with disseminated MAC (DMAC) infection, nine patients died during hospitalization . Another three cases died within a few months of appropriate anti-MAC chemotherapy . We concluded that the risk of DMAC infection in advanced AIDS patients in Thailand is high when low CD4 lymphocyte count is established . The prolonged fever resulted from DMAC in advanced HIV infection is warrant to be public health concern . Mycobacterium blood culture is a most valuable tool contributing to the diagnosis of infectious agents in this condition . The guidelines of 1997 USPHS/IDSA should be followed to give chemoprophylaxis against DMAC disease in patients with advanced HIV infection and a CD4 count less than 50 cells/mm3.

Clin Diagn Lab Immunol, 1998 Nov, 5(6), 826 - 30
Use of the 27-kilodalton recombinant protein from Paracoccidioides brasiliensis in serodiagnosis of paracoccidioidomycosis; Ortiz BL et al.; Paracoccidioidomycosis (PCM) is one of the most important endemic mycoses in Latin America; it is usually diagnosed by observation and/or isolation of the etiologic agent, Paracoccidioides brasiliensis, as well as by a variety of immunological methods . Although the latter are effective, two circumstances, cross-reactions with other mycotic agents and antigen preparation that is marked by extreme variability among lots, hinder proper standardization of the procedures . To circumvent this lack of reproducibility, molecular biology tools were used to produce a recombinant 27-kDa-molecular-mass antigen from this fungus; a sizable quantity of this antigen was obtained through fermentation of Escherichia coli DH5alpha, which is capable of expressing the fungal protein . The latter was purified by the Prep-Cell System (Bio-Rad); the recovery rate of the pure protein was approximately 6% . A battery of 160 human serum samples, consisting of 64 specimens taken at the time of diagnosis from patients with PCM representing the various clinical forms plus 15 serum specimens each from patients with histoplasmosis and aspergillosis, 10 each from patients with cryptococcosis and tuberculosis, 6 from patients with coccidioidomycosis, and 40 from healthy subjects, were all tested by an indirect enzyme-linked immunosorbent assay with the purified 27-kDa recombinant protein . The latter was used at a concentration of 1.0 microgram/well; there were three serum dilutions (1:1,000, 1:2,000, and 1:4,000) . The experiment was repeated at least twice . The average sensitivity for both experiments was 73.4%; in comparison with the healthy subjects, the specificity for PCM patients was 87.5% while for patients with other mycoses, it was 58.7% . Important cross-reactions with sera from patients with aspergillosis and histoplasmosis were detected . The positive predictive value of the test was 90.4% . These results indicate that it is possible to employ recombinant antigenic proteins for the immunologic diagnosis of PCM and, by so doing, achieve high coverage rates . Furthermore, antigen reproducibility can now be ensured, thus facilitating inter- and intralaboratory standardization.

Clin Diagn Lab Immunol, 1998 Nov, 5(6), 823 - 5
Improved immunodiagnosis of human candidiasis by an enzyme-linked immunosorbent assay using a Candida albicans 52-kilodalton metallopeptidase; El Moudni B et al.; An immunogenic aminopeptidase of Candida albicans was purified by high-performance liquid chromatography . It was then used for the development of an enzyme-linked immunosorbent assay to detect antibodies directed against this antigen in sera from patients with candidiasis . This enzyme specifically cleaves the L-Arg-7-amino-4-methyl-coumarin substrate at pH 7.4 and was detected in the crude extract of different C . albicans isolates . Sera used for this study were obtained from healthy blood donors or from patients with one of the following: systemic candidiasis, aspergillosis, cryptococcosis, toxoplasmosis, or malaria . The statistical analysis demonstrates significant differences between absorbency values obtained with sera from patients with candidiasis and with sera from the other groups (P = 0.000001) . Diagnostic parameters show high diagnostic specificity of 97% and a sensitivity of 83% at a cutoff value of 0.425 and suggest the usefulness of this aminopeptidase for the diagnosis of systemic candidiasis.

Arch Microbiol, 1998 Nov, 170(6), 395 - 404
Signaling via cAMP in fungi: interconnections with mitogen-activated protein kinase pathways; Kronstad J et al.; The cAMP signal transduction pathway controls a wide variety of processes in fungi . For example, considerable progress has been made in describing the involvement of cAMP pathway components in the control of morphogenesis in Saccharomyces cerevisiae, Ustilago maydis, and Magnaporthe grisea . These morphological processes include the establishment of filamentous growth in S . cerevisiae and U . maydis, and the differentiation of an appressorial infection structure in M . grisea . The discovery that appressorium formation requires cAMP signaling provides an immediate connection to fungal virulence . This connection may have broader implications among fungal pathogens because recent work indicates that cAMP signaling controls the expression of virulence traits in the human pathogen Cryptococcus neoformans . In this fungus, cAMP also influences mating, as has been found for Schizosaccharomyces pombe and as may occur in U . maydis . Finally, cAMP and mitogen-activated protein kinase pathways appear to function coordinately to control the response of certain fungi, e.g., Saccharomyces cerevisiae and Schizosaccharomyces pombe, to environmental stress . There are clues that interconnections between these pathways may be common in the control of many fungal processes.

J Thorac Imaging, 1998 Oct, 13(4), 247 - 60
Approach to the diagnosis of pulmonary disease in patients infected with the human immunodeficiency virus; Haramati LB et al.; Patients infected with the human immunodeficiency virus are predisposed to develop a variety of common and uncommon infectious and neoplastic pulmonary diseases . Clinical information that can stratify the risk of occurrence of these pulmonary conditions includes: 1) CD4 cell count-the most important determinant; 2) concurrent antimicrobial therapy; 3) prior travel history; 4) known latent infections that may reactivate: and 5) underlying respiratory disease . Specific pulmonary diseases are discussed including: bacterial pneumonia, bronchitis, mycobacterial and fungal infections, pneumocystis carinii pneumonia, toxoplasmosis, cytomegalovirus, Kaposi sarcoma, lymphoma, and lung cancer . A differential diagnosis can be generated based on the chest radiographic pattern . Focal or multifocal areas of consolidation usually represent conventional bacterial pneumonia or, less commonly, tuberculosis . In severely immunocompromised patients, unusual diseases causing consolidation should be considered including: Rhodococcus infection, nocardiosis, cryptococcosis, aspergillosis, and lymphoma . Nodules can be present in tuberculosis, histoplasmosis, cryptococcosis, and Kaposi sarcoma . Interstitial opacities are common in pneumocystis carinii pneumonia, histoplasmosis, and cytomegalovirus pneumonia . Cavitation and cysts are features of pneumocystis carinii pneumonia, tuberculosis, aspergillosis, and lung cancer . Disease of the airways is increasingly recognized in those with acquired immunodeficiency syndrome . Lymphadenopathy is most common in mycobacterial infection, but can be a feature of fungal infection, lymphoma, Kaposi sarcoma, and lung cancer . The combined use of clinical information, knowledge of typical conditions associated with the human immunodeficiency syndrome, and radiographic patterns offers a useful approach to the diagnosis of pulmonary disease in the patient with the human immunodeficiency virus.

Thorax, 1998 Jul, 53(7), 554 - 7
Cryptococcosis in HIV negative patients: findings on chest radiography; Roebuck DJ et al.; BACKGROUND: The findings at chest radiography in HIV negative patients with cryptococcosis are said to differ from those in HIV positive patients . No large series of HIV negative patients has been reported . METHODS: A review of the medical charts and radiographs of 44 consecutive HIV negative patients with proven cryptococcosis was undertaken . RESULTS: Air space consolidation involving one or more lobes was found in 15 patients (34%), one or more pulmonary masses in 13 patients (30%), and interstitial opacities in three patients (7%) . No other radiographic finding was detected in more than one patient . Twelve of 23 patients (52%) with cryptococcal meningitis had abnormal chest radiographs, and 10 of 16 patients (62%) who presented with respiratory symptoms had air space consolidation . CONCLUSIONS: HIV negative patients tend to have different radiographic findings from those reported in patients with AIDS . Cryptococcal meningitis in this group is usually associated with pulmonary mass(es) or a normal chest radiograph, whereas a respiratory presentation is usually associated with air space consolidation.

Antimicrob Agents Chemother, 1998 Nov, 42(11), 2961 - 5
Specific activities of dolastatin 10 and peptide derivatives against Cryptococcus neoformans; Pettit RK et al.; The biosynthetic peptide dolastatin 10 is currently in phase I and II cancer clinical trials . We evaluated the antifungal spectrum of dolastatin 10 and four structural modifications . In broth macrodilution assays, the peptides were fungicidal for American Type Culture Collection strains and clinical isolates (including fluconazole-resistant strains) of Cryptococcus neoformans but no other yeasts or filamentous fungi examined . Specificity for C . neoformans was also demonstrated in the solid-phase disk diffusion assay, and fungicidal activity was confirmed in time-kill experiments . For a methyl ester modification, the MICs at which 50 and 90% of 19 clinical isolates were inhibited (MIC50 and MIC90, respectively) were 0.195 and 0.39 microg/ml, respectively . The MFC50 (50% minimum fungicidal concentration) for this peptide was 0.39 microg/ml, and the MFC90 was 0.78 microg/ml . MICs and MFCs were identical or lower in the presence of human serum but increased with lowered pH . These peptides should be pursued as potential chemotherapeutics for C . neoformans, a leading cause of infection and mortality in immunocompromised patients.

Antimicrob Agents Chemother, 1998 Nov, 42(11), 2863 - 9
Sordarins: in vitro activities of new antifungal derivatives against pathogenic yeasts, Pneumocystis carinii, and filamentous fungi; Herreros E et al.; GM 193663, GM 211676, GM 222712, and GM 237354 are new semisynthetic derivatives of the sordarin class . The in vitro antifungal activities of GM 193663, GM 211676, GM 222712, and GM 237354 against 111 clinical yeast isolates of Candida albicans, Candida kefyr, Candida glabrata, Candida parapsilosis, Candida krusei, and Cryptococcus neoformans were compared . The in vitro activities of some of these compounds against Pneumocystis carinii, 20 isolates each of Aspergillus fumigatus and Aspergillus flavus, and 30 isolates of emerging less-common mold pathogens and dermatophytes were also compared . The MICs of GM 193663, GM 211676, GM 222712, and GM 237354 at which 90% of the isolates were inhibited (MIC90s) were 0.03, 0.03, 0.004, and 0.015 microg/ml, respectively, for C . albicans, including strains with decreased susceptibility to fluconazole; 0.5, 0.5, 0.06, and 0.12 microg/ml, respectively, for C . tropicalis; and 0.004, 0.015, 0.008, and 0.03 microg/ml, respectively, for C . kefyr . GM 222712 and GM 237354 were the most active compounds against C . glabrata, C . parapsilosis, and Cryptococcus neoformans . Against C . glabrata and C . parapsilosis, the MIC90s of GM 222712 and GM 237354 were 0.5 and 4 microg/ml and 1 and 16 microg/ml, respectively . The MIC90s of GM 222712 and GM 237354 against Cryptococcus neoformans were 0.5 and 0.25 microg/ml, respectively . GM 193663, GM 211676, GM 222712, and GM 237354 were extremely active against P . carinii . The efficacies of sordarin derivatives against this organism were determined by measuring the inhibition of the uptake and incorporation of radiolabelled methionine into newly synthesized proteins . All compounds tested showed 50% inhibitory concentrations of <0.008 microg/ml . Against A . flavus and A . fumigatus, the MIC90s of GM 222712 and GM 237354 were 1 and 32 microg/ml and 32 and >64 microg/ml, respectively . In addition, GM 237354 was tested against the most important emerging fungal pathogens which affect immunocompromised patients . Cladosporium carrioni, Pseudallescheria boydii, and the yeast-like fungi Blastoschizomyces capitatus and Geotrichum clavatum were the most susceptible of the fungi to GM 237354, with MICs ranging from </=0.25 to 2 microg/ml . The MICs of GM 237354 against Trichosporon beigelii and the zygomycetes Absidia corymbifera, Cunninghamella bertholletiae, and Rhizopus arrhizus ranged from </=0.25 to 8 microg/ml . Against dermatophytes, GM 237354 MICs were >/=2 microg/ml . In summary, we concluded that some sordarin derivatives, such as GM 222712 and GM 237354, showed excellent in vitro activities against a wide range of pathogenic fungi, including Candida spp., Cryptococcus neoformans, P . carinii, and some filamentous fungi and emerging invasive fungal pathogens.

P N G Med J, 1996 Sep, 39(3), 196 - 9
Ocular manifestations of AIDS; Verma N et al.; The acquired immune deficiency syndrome (AIDS) is a lethal multisystem disease . Its ocular manifestations have received relatively little attention in the literature . Between 73% and 100% of AIDS patients develop ocular lesions . The commonest lesions seen are retinal--either infectious or noninfectious retinopathy . Involvement of the conjunctiva with Kaposi's sarcoma, infected tears and infected cornea as well as the vitreous are less common . Infections with cytomegalovirus and varicella zoster virus are common causes of visual loss and can be treated with antiviral agents such as ganciclovir and foscarnet . This greatly increases the quality of life in these patients by preventing visual lossPIP: Ocular manifestations of AIDS, which occur in 73-100% of AIDS patients, are important to diagnose given their treatability . The most common AIDS-related ocular lesions are cotton wool spots and noninfectious retinopathies, cytomegalovirus retinitis, and conjunctival Kaposi's sarcoma . Less common are herpes zoster ophthalmicus, retinal toxoplasmosis, choroidal Pneumocystis carinii infection, herpes simplex and herpes zoster retinitis, and cryptococcal choroiditis . Administration of antiretroviral agents such as ganciclovir and foscarnet can prevent blindness, but their serious toxicity and the lack of oral preparations remain problematic .

P N G Med J, 1996 Sep, 39(3), 174 - 80
HIV infection and AIDS; Lloyd A; Many of the clinical features of HIV/AIDS can be ascribed to the profound immune deficiency which develops in infected patients . The destruction of the immune system by the virus results in opportunistic infection, as well as an increased risk of autoimmune disease and malignancy . In addition, disease manifestations related to the virus itself may occur . For example, during the primary illness which occurs within weeks after first exposure to HIV, clinical symptoms occur in at least 50% of cases, typically as a mononucleosis syndrome . HIV-related complications are rarely encountered in patients with preserved immunity (i.e . CD4 T-cell counts greater than 500 cells/mm3) . Recurrent mucocutaneous herpes simplex (HSV), herpes zoster (VZV), oral candidiasis and oral hairy leukoplakia occur with increasing frequency as the CD4 count drops below this level . Immune thrombocytopenia (ITP) occurs in association with HIV and often presents early in the clinical course . The risk of developing opportunistic infections and malignancies typical of AIDS increases progressively as CD4 counts fall below 200 cells/mm3 . The clinical manifestations of infections associated with AIDS tend to fall into well-recognized patterns of presentation, including pneumonia, dysphagia/odynophagia, diarrhoea, neurological symptoms, fever, wasting, anaemia and visual loss . The commonest pathogens include Candida albicans, Pneumocystis carinii, Mycobacterium tuberculosis, Toxoplasma gondii, Cryptococcus neoformans, Mycobacterium avium intracellulare and cytomegalovirus . Malignant disease in patients with HIV infection also occurs in a characteristic pattern . Only two tumours are prevalent: Kaposi's sarcoma, a multifocal tumour of vascular endothelium which typically involves skin and mucosal surfaces; and non-Hodgkin's lymphoma, which is typically high grade in phenotype, often arising within the central nervous system . The principles of therapy include reduction of HIV replication by antiretroviral agents, prophylaxis against the common opportunistic infections and treatment followed by subsequent lifelong maintenance therapy for infections when they do occurPIP: This article presents basic information on the clinical features of HIV infection, most of which are related to the profound immune deficiency associated with HIV/AIDS . Primary HIV infection is associated with clinical symptoms, primarily a mononucleosis syndrome, in about 50% of cases . In the ensuing 10 years, more than 50% of HIV-infected individuals develop the opportunistic infections (OIs) indicative of the onset of AIDS . Common presentations of AIDS include pneumonia, dysphagia, diarrhea, neurologic symptoms, fever, wasting, anemia, and vision loss . Monitoring of peripheral blood CD4 T-lymphocytes provides a measure of the current risk of OIs and a guide for antiretroviral therapy . Protease inhibitors, used in combination with other antiretrovirals, allow long-term control of HIV disease, but the substantial cost of these drugs has prohibited their widespread use in developing countries . Treatment of HIV-related infections must be followed by a maintenance regimen intended to prevent relapse .

J Formos Med Assoc, 1998 Sep, 97(9), 642 - 5
False-positive 2-{F-18}-fluoro-2-deoxy-D-glucose positron emission tomography studies for evaluation of focal pulmonary abnormalities; Yen RF et al.; Positron emission tomography (PET) with 2-{F-18}-fluoro-2-deoxy-D-glucose (FDG) can demonstrate the glucose metabolism characteristics of a lesion, which may be helpful in differentiating between benign and malignant focal pulmonary lesions . Malignant cells demonstrate higher glucose metabolic activity than benign lesions . However, some inflammatory processes also show significant FDG uptake . We present two cases where high FDG uptake was found in inflammatory lesions in the lungs . The first case was that of a 38-year-old woman with chronic cough for more than 20 years . FDG PET revealed a hypermetabolic lesion with a lesion-to-background ratio of 8.0 at the posterior aspect of the right middle lung . She underwent thoracotomy and tumor resection, and was diagnosed with cryptococcosis . The second case was that of a 72-year-old woman who had pulmonary tuberculosis previously with cavitation in the left lower lobe . She suffered from fever, chills and severe hemoptysis for several days before this admission . FDG PET revealed a hypermetabolic ring at the periphery of the cavity . The lesion-to-background ratio was 7.8 . Echo-guided biopsy showed no evidence of malignancy . She was treated with antibiotics and the symptoms subsided gradually . Lung abscess complicating a pre-existing cavity was diagnosed . These two cases substantiate that positive FDG PET results should be interpreted with caution in differentiating benign from malignant pulmonary abnormalities, especially in regions with a high prevalence of granulomatous lesions.

Nippon Ishinkin Gakkai Zasshi, 1998, 39(4), 225 - 8
{Mucocutaneous fungal infection in AIDS patients}; Ishii N; Human immunodeficiency virus (HIV) infection is unique among the cutaneous fungal infections caused by defects of the cell-mediated immune system . A complex interplay exists in the host between fungal virulence factors favoring disease, and immune and non-immune host mechanisms defending against disease . Cell-mediated immunity appears of paramount importance in defense against histoplasmosis, cryptococcosis, mucocutaneous candidiasis, and dermatophytosis . Mucocutaneous candidiasis is especially common in patients with HIV infection . Anti-fungal medicines were effective against candidiasis . However, anti-fungal medicine-resistant candida has been observed . Anti-fungal ointment was effective against dermatophytosis and seborrheic dermatitis . No cases of cutaneous fungal infection resistant to anti-fungal medicine have been observed.

Nippon Ishinkin Gakkai Zasshi, 1998, 39(4), 193 - 7
{Severe mycosis-factors with fungus and host}; Maesaki S et al.; The deep-seated mycosis occurred in immunocompromized patients . Normally, the deep-seated mycosis became sever infection because of the defects of host defense or the adverse effects of antifungal agents . The major factors of the reason on the severity of the deep-seated mycosis depends on the pathogenesity and the drug resistance for antifungal agents in infected fungi . The clinical factors related with hosts defenses are important to the other reason on the severity . We investigated that the multiple drug resistant (MDR) mechanism may be one of the major roles plays in the azole resistant Candida albicans strains isolated form the patients with oropharyngeal candidiasis infected HIV . We analyzed which clinical factors are related with the prognosis of the patients with pulmonary cryptococcosis and aspergilloma . The titer of cryptococcal capsular antigen was earlier improve in the patient without underline disease than in the patients with underline diseases diagnosed pulmonary cryptococcosis . CRP was higher in the death cases in the patients with pulmonary aspergilloma, compared with alive cases.

Dtsch Tierarztl Wochenschr, 1998 Sep, 105(9), 349 - 53
{Distribution, serovar affiliation and epidemiologic behavior of Cryptococcus neoformans isolates from ornamental bird breeds}; Kielstein P et al.; Investigations of faeces samples from breeding stocks of companion birds in the federal state of Thuringia revealed a high contamination rate of companion birds with Cryptococcus (Cr.) neoformans var . neoformans . The prevalence of Cr . neoformans var . neoformans correlated with the spectrum of bird species present in the respective breeding units . The causes for that are not clear at the moment . Sensitivity of Cr . neoformans var . neoformans towards alkaline agents was not confirmed and was ruled out as a reason for different tenacity of the yeast in various bird breedings . Differentiation of varieties within Cr . neoformans was possible on the basis of proline assimilation, determination of canavanine resistance, EDTA urease test, as well as Cr . neoformans var . neoformans factor sera and PCR fingerprinting . Serological differentiation of serovars and PCR fingerprinting resulted in subdivision of Cr . neoformans var . neoformans isolates into two groups, which corresponded to serovars A and D . A prevalence of serovar A isolates was found in investigated bird breeding stocks . This also corresponded to the distribution of Cr . neoformans var . neoformans described in literature in humans with cryptococcosis in Germany . Consequently, serovar A or D infections of patients may be connected with their contacts to Cr . neoformans-excreting companion birds.

AIDS, 1998 Oct 1, 12(14), 1815 - 22
Opportunistic infections occurring during highly active antiretroviral treatment; Michelet C et al.; OBJECTIVE: To analyse the characteristics of opportunistic infections in patients receiving highly active antiretroviral treatment (HAART) . DESIGN AND METHODS: A retrospective study performed in seven hospitals, included all patients starting treatment by ritonavir or indinavir between 26 March and 31 December 1996 . Patients were evaluated for the development of AIDS-defining events . Clinical evaluation, plasma HIV-1 RNA quantification, CD4 cell count were recorded at baseline and at the onset of the event . RESULTS: Four hundred and eighty-six patients were included: 44.2% had a CD4 cell count below 50 x 10(6) cells/l . Fifty clinical events were recorded in 46 patients with a mean follow-up of 6.1 months, of which 34 events (68%) were observed during the first 2 months of HAART . Eighteen of these occurred despite a reduction of viral load by at least 1.5 log10) and a 100% increase of the CD4 cell count compared with that at the onset of the event, corresponding to 11 cytomegalovirus infections, five mycobacterial infections, one case of cryptococcosis, and one case of Varicella-Zoster virus-related acute retinal necrosis . Among the 16 events observed after the second month, six occurred despite a marked biological improvement, corresponding to a recurrence in five of six patients who had stopped their maintenance therapy . Events were one cytomegalovirus infection, two mycobacterial infections, one episode of oesophageal candidiasis and one cryptococcal meningitis . CONCLUSION: In patients at high risk of developing an opportunistic infection prior to the institution of a HAART regimen, prophylaxis should not be discontinued during the first 2 months of treatment, and maintenance therapy should be carried on despite a significant increase in the CD4 cell count.

Cell Immunol, 1998 Nov 1, 189(2), 144 - 8
Development of macrophage anticryptococcal activity in vitro is dependent on endogenous M-CSF; Brummer E et al.; We previously showed that nonactivated resident murine peritoneal macrophages (PM) from five strains (e.g., BALB/c) have C'-dependent fungistasis for Cryptococcus neoformans in 24-h coculture, but not CD-1 PM unless culture time was extended or M-CSF treatment was used . We studied effect of a rat IgG1 monoclonal (m) antibody (Ab) to murine M-CSF receptor on this anticryptococal activity . Culture of BALB/c PM with mAb, diluted 1:10, prechallenge reduced fungistasis from 58 to 21% (P < 0.01), whereas further 10-fold dilutions did not . Moreover, M-CSF pretreatment (5000 U/ml) significantly enhanced fungistasis (to 85%), whereas adding mAb 1:10 or 1:100 reduced that (to 58 and 77%, respectively, P < 0.01) . In 48-h culture CD-1 PM had 39% fungistasis, reduced to 0% by mAb, M-CSF treatment of CD-1 PM increased fungistasis to 72%, which was reduced to 13 or 58% (P < 0 . 001) by 1:10 or 1:100 mAb, respectively . Complete blocking by mAb of CD-1 PM activity was consistent with lack of measurable early endogenous CD-1 M-CSF production . Increasing exogenous M-CSF could overcome the inhibition by mAb (64% fungistasis BALB/c PM reduced to 11% with inhibition by mAb or increased to 94% with 5000 U/ml M-CSF; 37% with both mAb and M-CSF, 51% with mAb and 10,000 U/ml; P < 0.05, 5000 U/ml + mAb vs 10,000 U/ml + mAb) . Moreover, rabbit Ab to M-CSF significantly reduced anticryptococcal activity of untreated BALB/c macrophages . In summary, development of PM fugistatic activity is dependent on endogenous M-CSF, since it is blocked by anti-receptor mAb (as is exogenous M-CSF stimulation) or anti-M-CSF Ab, and macrophages of the mouse strain with delayed activity had no measurable early M-CSF production .

Rev Soc Bras Med Trop, 1998 Sep-Oct, 31(5), 465 - 72
{Renal anatomopathological changes in patients with acquired immunodeficiency deficiency syndrome}; Laguna-Torres VA et al.; Renal anatomopathological lesions were studied among 119 AIDS patients from Faculdade de Medicina do Triangulo Mineiro's University Hospital (Uberaba, MG, Brazil) . From formalin-fixed blocks, slides were obtained and studied by light microscopy . Of 119 patients, 67 presented tubulointerstitial nephritis (TIN), 18 inespecific, 2 xantogranulomatous and infections agents were found in 48 as follows: mycosis in 28 (16 Cryptococcus sp; 9 Histoplasma sp, 1 Candida sp e 2 Paracoccidioides brasiliensis); bacteria in 18 (9 Mycobacterium sp), virus in 6 (Cytomegalovirus) . Acute tubular necrosis was found in 43 cases (36.1%) . Other diagnosis were: nefrocalcinosis (15.1%), arteriolar hyalinosis (22.7%), two cases of focal segmental glomerulosclerosis (1.7%) and one case of diffuse mesangial hyperplasia (0.8%) . We conclude that the renal involvement in patients with AIDS, presents a wide spectrum of pathologies, secondary to complications related to opportunistic infections, therapeutic and diagnostic management, and the nephropathies associated to HIV.

J Antimicrob Chemother, 1998 Sep, 42(3), 321 - 31
Multi-centre evaluation of the Etest method for antifungal drug susceptibility testing of Candida spp . and Cryptococcus neoformans . BSAC Working Party on Antifungal Chemotherapy; Warnock DW et al.; Ten laboratories tested 18 isolates of Candida spp . and two of Cryptococcus neoformans against amphotericin B, flucytosine, fluconazole and itraconazole on two occasions by the Etest method . Two individuals read each set of results . Of the 18 isolates of Candida spp., five were duplicated, but the participants were not told this . In 40 of the 60 drug-organism combinations studied, at least 80% of the Etest MICs fell within a five-concentration range corresponding to the modal MIC +/- 1 log2 dilution . In five combinations, >50% of the Etest MICs fell outside this five-concentration range . In 17 (85%) of the 20 drug-organism combinations tested in duplicate, at least 80% of the paired Etest results fell within two concentrations of each other (corresponding to one log2 dilution) . Overall, 88.5% of the paired Etest results for amphotericin B agreed to within two concentrations, as did 94% of results for flucytosine, 92% for fluconazole and 79% for itraconazole . The broth microdilution MICs of the four antifungal agents for the 15 isolates were measured on five occasions in the Mycology Reference Laboratory, Bristol . In each case, the results fell within a three log2 concentration range . In 24 (40%) of the 60 drug-organism combinations tested, at least 80% of the Etest results fell within the broth microdilution test MIC range, but 27 (45%) showed <50% exact agreement . In 33 (73%) of 45 drug-organism combinations involving flucytosine, fluconazole or itraconazole, at least 80% of the Etest results fell within the same class (susceptible, resistant, or susceptible dependent upon dose) as the broth microdilution results . With fluconazole, the Etest method misclassified three susceptible isolates of Candida spp . as resistant in 1.5-15% of tests . With itraconazole, the Etest misclassified seven susceptible isolates of Candida spp . as resistant in 5-62.5% of tests . The Etest also misclassified both C . neoformans isolates as resistant to flucytosine, fluconazole and itraconazole in 7.5-65% of tests . Our results suggest that the Etest is suitable for routine use with Candida spp . and amphotericin B or flucytosine . It is less reliable for the azoles, and isolates that appear to demonstrate acquired resistance should be retested with well-established reference methods.

Neurology, 1998 Oct, 51(4), 1213 - 5
Dual infective pathology in patients with cryptococcal meningitis; Silber E et al.; Coinfection of the nervous system by two distinct nonviral organisms is uncommon and often undiagnosed . Medical teaching emphasizes that a single pathologic process should be sought; however, in the presence of severe immunocompromise this approach may not hold true . We describe seven HIV-1 seropositive patients with cryptococcal meningitis, three of whom had a proven nervous system infection with a second organism: concurrent tuberculous meningitis, a tuberculoma, and the first documented case of cryptococcal meningitis and neurosyphilis.

J Infect Dis, 1998 Nov, 178(5), 1464 - 71
Cryptococcus neoformans and Candida albicans regulate CD4 expression on human monocytes; Pietrella D et al.; This study examined the capability of Candida albicans and Cryptococcus neoformans to modulate CD4 expression on human monocytes . C . albicans and an acapsular strain of C . neoformans induced higher levels of CD4 expression than encapsulated strains . Purified glucuronoxylomannan did not regulate CD4 expression on monocytes, but down-regulation of CD4 expression compared with stimulation by acapsular C . neoformans alone was observed when glucuronoxylomannan was used in combination with acapsular C . neoformans . The ability of opsonic factors to facilitate fungal-mediated CD4 overexpression suggests that binding or internalization (or both) of the yeast cells is a critical event . Protein synthesis was required, excluding redistribution of the intracellular pool of CD4 receptors to the cellular surface as the sole possible mechanism . Results demonstrate a new effect of fungi on professional phagocytic cells and raise the possibility that modulation of CD4 could influence gp120-mediated human immunodeficiency virus entry.

Pathol Res Pract, 1998, 194(8), 567 - 70
AIDS-associated nephropathy: 5-year retrospective morphologic analysis of 87 cases; Soriano-Rosas J et al.; Morphologic findings in the kidneys of 138 consecutive acquired immunodeficiency syndrome (AIDS) autopsies performed at the Mexico City General Hospital between 1986 and 1991 were studied . A total of 87 cases (63%) had renal disease, 62 cases presented glomerular alterations, and 80 cases showed tubulointerstitial damage . Glomerular collapse was seen in 30 cases (48%), mesangial expansion in 22 cases (35%), focal and segmental glomerulosclerosis in 20 cases (32%) and glomerular proliferation in only eight cases (11%) . Nephrocalcinosis was seen in 25 cases (31%) . Thirty-six cases (45%) presented tubulointerstitial infections: Mycobacterium tuberculosis in 19 cases (23%), Cryptococcus in 10 cases (12.5%), Cytomegalovirus CMV in eight cases (10%), Gram-negative bacteria in 3 cases (3.7%), and one case with histoplasmosis . In six cases the pathogens were multiple . Two cases showed infiltration of large-cell non-Hodgkin lymphoma . Based on the results obtained by autopsies, we conclude that in our country there is a high frequency of renal affection in AIDS patients . Clinically, this disease is infrequently diagnosed and not always related to the cause of death.

Drugs, 1998 Sep, 56(3), 365 - 83
Amphotericin-B colloidal dispersion . A review of its use against systemic fungal infections and visceral leishmaniasis; Brogden RN et al.; Formulation of amphotericin B with sodium cholesteryl sulphate alters the pharmacokinetic properties of the drug, particularly reducing its distribution to the kidneys . The antifungal activity in vitro of amphotericin B colloidal dispersion (ABCD) is similar to that of conventional amphotericin B (C-AmB) against true pathogenic organisms including Blastomyces, Coccidioides, Histoplasma and Paracoccidioides species and the opportunistic organisms such as Candida and Cryptococcus species . In animal models, ABCD was generally less effective than an identical dose of C-AmB, but overall was more effective because of its improved therapeutic index . Although ABCD appeared to be more effective than C-AmB in resolving infection and improving survival in patients with proven or probable invasive aspergillosis, the retrospective design of the study and the greater prevalence of neutropenia in patients treated with the conventional formulation necessitate cautious interpretation of the results . ABCD has been effective and seldom caused nephrotoxicity in patients with fungal infection who had previously failed to adequately respond or had developed renal toxicity with C-AmB . Similarly, ABCD was effective in patients with proven or suspected fungal infection after bone marrow transplantation . Preliminary results from a pilot study comparing ABCD and C-AmB in patients with neutropenia and persistent fever reported similar response rates with both formulations . ABCD is an effective treatment for visceral leishmaniasis in immunocompetent patients . In 1 study, about 12% of ABCD recipients discontinued the drug because of adverse events; infusion-related events were the most common cause of discontinuation . The renal tolerability of ABCD is better than that of C-AmB . ABCD appears to be an effective alternative to conventional amphotericin B in patients with invasive aspergillosis or visceral leishmaniasis and in those with proven or suspected systemic fungal infection who are intolerant of the conventional formulation or have pre-existing renal impairment . Preliminary data also suggest that ABCD is an alternative to C-AmB when used empirically in patients with neutropenia and fever . Nevertheless, the efficacy of ABCD compared with that of the conventional formulation has yet to be adequately demonstrated and the role of ABCD relative to that of liposomal and other lipid-based formulations has not been determined . CONCLUSIONS: ABCD, like other lipid-based and liposomal formulations of amphotericin B, has been designed to deliver the active drug to the target site, while reducing renal toxicity . The aim of increasing the therapeutic index compared with C-AmB has been achieved.

Med Mycol, 1998 Aug, 36(4), 227 - 33
Damage to yeast cells of Cryptococcus neoformans by voriconazole and fluconazole: a culture and microscopic study; Brummer E et al.; A systematic cultural, cytological and microscopic study of voriconazole (VCZ) and fluconazole (FCZ) damage to Cryptococcus neoformans over time was made . When haemocytometer counts were compared with colony-forming units (cfu) viability decreased with increased drug concentration and prolonged treatment time up to 48 h . Percentage viability by vital staining correlated with cfu . Concentrations of VCZ were found to be 10-fold more potent than FCZ . At 72 h, percentage viability increased in cultures with lower drug concentrations, indicating outgrowth of surviving yeast cells . Drug treatment resulted in a cytological change in a large percentage of yeast cells characterized by a large central vacuole easily observed microscopically . Vital staining showed that there was no direct relationship between cytological changes and non-viability . These novel findings add a new approach for studying the antifungal action of VCZ and FCZ against C . neoformans and provide a new perspective on their antifungal action.

Med Mycol, 1998 Aug, 36(4), 189 - 97
Role of the capsule in microglial cell-Cryptococcus neoformans interaction: impairment of antifungal activity but not of secretory functions; Barluzzi R et al.; Using two isogenic strains of Cryptococcus neoformans, we studied the influence of the capsule in C . neoformans microglial-cell interaction . We demonstrate that the acapsular mutant yeasts (CAP67) are more susceptible to phagocytosis and killing than encapsulated yeasts (B3501) by the murine microglial cells, BV-2 . RT-PCR analysis showed that the pattern of gene transcripts for tumour necrosis factor alpha (TNF-a), interleukin (IL)-1beta, IL-6, IL-12p40 and granulocyte macrophage colony stimulating factor remains unchanged following BV-2 cell infection with CAP67 or B3501 yeasts . Moreover, no induction of TNF-alpha secretion occurs in BV-2 cells infected with either B3501 or CAP67 yeasts or exposed to glucuronoxylomannan (GXM) or galactoxylomannan (GalXM) . Nevertheless, lipopolysaccharide-induced TNF-alpha secretion is downregulated by cell infection with B3501 or CAP67 yeasts or exposure to GXM or GalXM . Overall, by means of a continuous cell line, it appears that the C . neoformans capsule is detrimental to microglial cell antifungal activity, while no effect can be attributed to the capsule as trend of cytokine gene expression and TNF-alpha secretion.

Med Mycol, 1998 Jun, 36(3), 185 - 8
Analysis of HLA association in susceptibility to infection with Cryptococcus neoformans var . gattii in a Papua New Guinean population; Van Dam MG et al.; The possible association between susceptibility to infection with Cryptococcus neoformans var . gattii and HLA phenotype was examined in a group of Papua New Guinean patients . There was no evidence for a statistically significant association between susceptibility to infection and HLA class I and HLA class II phenotypes, although analysis of data which had not been subjected to the appropriate Bonferroni correction factor suggested a trend for susceptibility linked to HLA B*5601.

Med Mycol, 1998 Jun, 36(3), 169 - 75
Specific identification of Penicillium marneffei by a polymerase chain reaction/hybridization technique; Vanittanakom N et al.; Penicillium marneffei has been described recently as a cause of an emerging mycotic infection in HIV-infected patients . A PCR/hybridization assay was developed to rapidly identify this pathogen . The nucleotide sequence of the 631-bp region of 18S ribosomal DNA of P . marneffei was determined using the standard dideoxy chain termination method . An oligonucleotide probe was designed on the basis of the analysed sequences of P . marneffei and 18S rDNA sequences of other fungi in the GenBank database . A 631-bp PCR product was amplified using primers RRF1 and RRH1 from P . marneffei and seven other fungi, Penicillium spp., Aspergillus fumigatus, A . flavus, Histoplasma capsulatum, Cryptococcus neoformans, Candida albicans and C . krusei . A 15 oligonucleotide segment (Pm3) which was specific for P . marneffei was synthesized and used as a probe . Only the PCR products of P . marneffei isolates hybridized with the Pm3 oligonucleotide probe . The sensitivity of the assay was approximately 0.5 pg/microl and 0.1 pg/microl of DNA by PCR and Southern hybridization, respectively . The usefulness of this method as a diagnostic tool will require further studies.

Med Mycol, 1998 Apr, 36(2), 119 - 22
Cryptococcus neoformans var . gattii--evidence for a natural habitat related to decaying wood in a pottery tree hollow; Lazera MS et al.; To study hollows of living trees as the natural habitat of Cryptococcus neoformans in an endemic area of cryptococcosis in the northeastern Brazilian region, samples of decaying wood were collected inside the hollows, plated on niger seed agar and inoculated into mice and hamsters . Identification of C . neoformans was based on morphological and physiological tests . Canavanine-glycine-bromothymol medium was used to screen the varieties and Crypto Check Iatron Kit to serotype the isolates . For a period of 29 months C . neoformans var . gattii serotype B was isolated repeatedly from the hollow of a pottery tree (Moquilea tomentosa), pointing to the natural occurrence of C . neoformans var . gatti in decaying wood forming hollows in living trees . Evidence for a natural habitat of the variety gattii other than that related to Eucalyptus camaldulensis are discussed.

Med Mycol, 1998 Feb, 36(1), 47 - 50
Simultaneous infection with Blastomyces dermatitidis and Cryptococcus neoformans; Mounts A et al.; In this report, we present two cases of simultaneous infection with Blastomyces dermatitidis and Cryptococcus neoformans . In both cases, fungi were isolated from the lungs . The clinical manifestations were not suggestive of infection with two fungi . One patient was receiving immunosuppressive therapy and another was otherwise immunocompetent . A review of the literature failed to uncover any reports of co-infection with these two fungal pathogens . We discuss possible mechanisms for the isolation of both pathogens.

Med Mycol, 1998 Feb, 36(1), 1 - 5
Interstrain variation in the deoxynucleotide composition of Cryptococcus neoformans: nucleotide composition of Cryptococcus neoformans; Podwall D et al.; The deoxynucleotide (dNMP) composition of ten strains of C . neoformans was analysed by 32P-labelling and two-dimensional thin-layer chromatography . This technique is very sensitive for detecting rare deoxynucleotide adducts and analogues (minor bases) in DNA . The results indicate considerable variation among strains in DNA nucleotide composition.

Clin Tech Small Anim Pract, 1998 Aug, 13(3), 167 - 78
Inflammatory diseases of the central nervous system in dogs; Thomas WB; Inflammatory diseases of the central nervous system (CNS) are important causes of seizures in dogs . Specific diseases include canine distemper, rabies, cryptococcosis, coccidioidomycosis, toxoplasmosis, neosporosis, Rocky Mountain spotted fever, ehrlichiosis, granulomatous meningoencephalomyelitis, and pug dog encephalitis . Inflammatory disorders should be considered when a dog with seizures has persistent neurological deficits, suffers an onset of seizures at less than 1 or greater than 5 years of age, or exhibits signs of systemic illness . A thorough history, examination, and analysis of cerebrospinal fluid are important in the diagnosis of inflammatory diseases . However, even with extensive diagnostic testing, a specific etiology is identified in less than two thirds of dogs with inflammatory diseases of the CNS.

J Clin Microbiol, 1998 Nov, 36(11), 3438 - 40
Development of a nested PCR for detection of Cryptococcus neoformans in cerebrospinal fluid; Rappelli P et al.; We report the development of a nested-PCR-based assay for the detection of Cryptococcus neoformans in cerebrospinal fluid . The specificity and sensitivity of the test were assessed . The technique was then applied to 40 cerebrospinal fluid samples . We obtained positive reactions for all 21 clinical samples from patients who had been previously diagnosed as having cryptococcal meningitis by conventional techniques and negative reactions for all 19 negative controls . Nested PCR is here compared with other diagnostic methods currently used in patients' follow-up exams during anticryptococcal therapy.

Pathology, 1998 Aug, 30(3), 316 - 7
Cryptococcal myositis: a case report and review of the literature; O'Neill KM et al.; Only rare cases of cryptococcal myositis have been previously reported in the literature . All of these cases have occurred in the setting of human immunodeficiency virus (HIV) infection . We report a case of cryptococcal myositis diagnosed premortem on a needle biopsy in a heart transplant patient undergoing immunosuppressive therapy.

Rev Cubana Med Trop, 1992, 44(2), 141 - 4
{Detection of anti-Cryptococcus neoformans antibodies in 3 groups of human sera}; Martinez Machin G et al.; The tube agglutination (TA) technique was normalized in order to determine anti-Cryptococcus neoformans antibodies, according to Palmer et al., by employing positive and negative control sera and an antigenic suspension prepared from an autochtonous strain of Cryptococcus neoformans varneofornnans . Three groups of human sera were studied and the role of TA in detecting antibodies in the group of sera from patients with active cryptococcosis (100%) was shown . In sera from banks positiveness was only 6%, while in the group of fowl breeders, considered to be "exposed", positiveness was found to be 16%, although with low titres . A discussion is carried out about the value of this technique as a complement in the diagnosis and prognosis of cryptococcosis.

Obstet Gynecol, 1998 Oct, 92(4 Pt 2), 682 - 4
Isolated pulmonary cryptococcosis in pregnancy; LaGatta MA et al.; BACKGROUND: Isolated pulmonary cryptococcosis in an immunocompetent parturient host is a rare event . Little is known about this condition, its prognosis, and its treatment in the immunocompetent pregnant woman . CASE: A 28-year-old woman, para 3, was diagnosed with isolated pulmonary cryptococcosis postpartum . She had delivered a healthy neonate at term who revealed no clinical features compatible with maternal-fetal transmission . The woman was monitored closely throughout the course of her disease and radiogic, clinical, and serologic resolution was achieved after treatment with fluconazole . CONCLUSION: Patients with pulmonary cryptococcosis present with clinical pictures similar to other well-known diseases such as septic pulmonary emboli or choriocarcinoma . Treatment with triazole antifungal therapy yielded favorable results in our patient with this rare condition.

Mo Med, 1998 Sep, 95(9), 539 - 41
Case report of long-term survival in a patient with acquired immunodeficiency syndrome and cryptococcal meningitis; Luchi M et al.; In the era before protease inhibitors were available, the great majority of patients with AIDS died within five years of the diagnosis . This grim reputation may cause both physician and patient to give up hope prematurely when antiretroviral therapy fails . We report a patient who survived five years after the diagnosis of cryptococcal meningitis and AIDS . Although there are now combinations of antiretroviral drugs available that can delay disease progression and extend the lives of AIDS patients, these are associated with a significant failure rate . It is thus important to be aware of the potential to extend life in patients even when antiretroviral therapy is not effective.

Am J Respir Cell Mol Biol, 1998 Oct, 19(4), 588 - 97
T and B cell independence of endothelial cell adhesion molecule expression in pulmonary granulomatous inflammation; Izzo AA et al.; A pulmonary Cryptococcus neoformans (Cne: strain 52D, ATCC24067) infection model in mice was used to examine the possible role for T cell-mediated immunity in regulating vascular adhesion molecules on lung endothelium during development of granulomatous inflammation . Resolution of pulmonary Cne infection in C.B-17 mice begins by Day 14 following intratracheal inoculation and depends on T cell-mediated recruitment of monocytes followed by their activation . C.B-17 scid/scid (SCID) mice mount a less exuberant pulmonary inflammatory response, recruit fewer monocytes into their lungs, and fail to clear the infection . Recruitment of leukocytes into infected tissue is mediated by both the interaction of adhesion molecules expressed on the surface of activated vascular endothelial cells with ligands on circulating cells, and the directed response of these leukocytes to chemotactic factors . The kinetics of expression of the endothelial cell adhesion molecules E-selectin, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1), all previously shown to regulate monocyte recruitment, were examined in the lungs of infected C.B-17 and SCID mice during pulmonary infection to determine if T cells were necessary for their upregulation . Immunohistochemical analysis showed that upregulation of E-selectin, VCAM-1, and ICAM-1 did not differ significantly between C.B-17 and SCID mice at any time during infection . Maximal expression in C.B-17 and SCID mice was noted between Days 5 and 7 for all three molecules and preceded maximal influx of leukocytes into the lung . Thus, the inability of SCID mice to recruit optimal numbers of monocytes into infected lungs was not the result of a failure to express the critical adhesion molecules early in infection, but likely reflected absence of immune dependent chemotactic factors.

Microbiol Res, 1998 Aug, 153(2), 137 - 43
Influence of temperature, pH and water activity on "in vitro" inhibition of Penicillium glabrum (Wehmer) Westling by yeasts; Sinigaglia M et al.; Four different yeast species (Metschnikowia pulcherrima, Saccharomycopsis vini, Kluyveromyces marxianus, Cryptococcus albidus), isolated from surface of grapes, were evaluated for biocontrol potential against Penicillium glabrum . In order to investigate the influence of temperature, pH, water activity and yeast cell concentration on Penicillium glabrum inhibition, the individual effects and the interaction of these factors were analyzed by means of a Central Composite Design (CCD) . All yeast species tested showed antagonistic effects which were more pronounced at high cell concentrations . The other variables affected the antagonistic effect differentially depending on the yeast species . Results of the experimental design showed that the selective success of a competitive microflora is under environmental control; moreover, when microbial cells are subjected to multiple factors, the effects and the reciprocal interactions of the individual variables cannot be independently evaluated.

J Immunol, 1998 Oct 1, 161(7), 3557 - 68
Monoclonal antibodies reveal additional epitopes of serotype D Cryptococcus neoformans capsular glucuronoxylomannan that elicit protective antibodies; Mukherjee J et al.; Epitope specificity and isotype influence mAb efficacy against Cryptococcus neoformans; however, the relative contribution of each attribute is poorly understood . To date, only mAbs that recognize two epitopes of capsular glucuronoxylomannan (GXM), defined by the IgG1 mAbs 2H1 and E1, consistently mediate protection against C . neoformans . The role of epitope specificity was further examined using six additional IgG1 mAbs and serotype D C . neoformans ATCC 24067 . mAbs 3C2, 439, and 471 recognize the 2H1 epitope, whereas mAbs 339, 1255, and 302 recognize two separate epitopes . mAbs 3C2, 439, and 471 competed for GXM with the IgA mAb 18G9, a 2H1 mAb family member, whereas mAbs 302, 339, and 1255 did not . Each mAb bound GXM similarly, as determined by agglutination, direct Ag binding, Ag inhibition, and indirect capsular immunofluorescence assays . mAb apparent affinity constants for GXM ranged from 5 to 26 x 10(7) M(-1) with mAb 1255 > 3C2 > 339 > 439 > 471 > 302 . Each mAb significantly prolonged survival (p < 0.05); the average survival times of control and mice passively immunized with mAbs 3C2, 302, 339, 439, 471, and 1255 were 10.8, 36.6, 33, 25.5, 24.9, 17, and 22.6 days, respectively . Although each mAb enhanced J774.16 cell fungicidal activity, differences were observed in the ability of each mAb to facilitate attachment and ingestion of cryptococci . These results indicate 1) two additional epitope specificities associated with mAb efficacy, 2) differences in opsonic and protective efficacy for IgG1 anti-GXM mAbs, 3) an association between affinity and protective efficacy, and 4) additional support for association between an annular indirect capsular immunofluorescence pattern and mAb efficacy.

Pharmacotherapy, 1998 Sep-Oct, 18(5), 1087 - 92
Case-control study of amphotericin B in a triglyceride fat emulsion versus conventional amphotericin B in patients with AIDS; Manfredi R et al.; We evaluated the efficacy and tolerability of amphotericin B triglyceride emulsion in 16 patients with acquired immunodeficiency syndrome-related candidal esophagitis and cryptococcosis, compared with standard amphotericin B in 24 patients . Compared with the conventional formulation, the fat emulsion was administered in a significantly greater daily dose, and required shorter induction period and infusion time (p<0.001-<0.03) . Although the two drugs had similar clinical and microbiologic efficacy, the fat emulsion had a better safety profile with respect to frequency of flu-like symptoms, other local and systemic adverse events, and treatment discontinuation (p<0.02-<0.05) . Because it is easily available and inexpensive, it may have a number of advantages over the conventional formulation . Further similar comparisons are warranted, in addition to investigations to assess whether reduced toxicity can be obtained with fat emulsion without impairing (or possibly improving) the efficacy of this key antifungal agent.

J Neurol, 1998 Sep, 245(9), 598 - 602
Diagnostic value of atypical lymphocytes in cerebrospinal fluid from adults with enteroviral meningitis; Sato Y et al.; We have noted two morphologically distinct types of atypical lymphocytes (AL) in the cerebrospinal fluid (CSF) of adult patients with meningitis: one, which we designate type-I AL, with multilobulated nuclei resembling those of the abnormal cells in adult T-cell leukaemia (ATL); and another, type-II AL, characterized by large lymphocytes with basophilic cytoplasm and nuclei containing coarse chromatin . Type-I AL were detected in 25 of 39 patients (64%) with enteroviral and in 11 of 109 (11%) with aseptic meningitis presumed to be caused by other viruses, but not in meningitis resulting from Cryptococcus neofirmans (n = 14), Mycobacterium tuberculosis (n = 19) or acute bacterial infection (n = 49) . Type-I AL were not seen in herpes zoster (n = 15) aseptic meningeal reactions (n = 15), or in leptomeningeal carcinomatosis (n = 14) . Type-II AL were often present in meningitis of various aetiologies and in aseptic meningeal reactions, but not in leptomeningeal carcinomatosis . The presence of type-I AL in the CSF was found to be indicative of enteroviral meningitis with the highest predictive value (69%), while type-II AL had a lower diagnostic positive predictive value in meningitis of the five aetiologies above . Type-I AL immunostained for CD4, while type-II AL were stained for CD8 . The presence of type-I AL in CSF strongly suggests enteroviral meningitis, which warrants careful follow-up without antifungal, antituberculous or antibacterial agents . However, type-I AL, which are likely to be virally transformed lymphocytes, must be distinguished from ATL cells, which frequently involve the meninges.

Int J Epidemiol, 1998 Aug, 27(4), 698 - 702
Causes of death in a rural, population-based human immunodeficiency virus type 1 (HIV-1) natural history cohort in Uganda; Okongo M et al.; BACKGROUND: While human immunodeficiency virus (HIV)-related causes of death have been well documented in developed countries, in Africa data are scanty and mainly based on autopsy studies from city hospitals which are highly selective and may not represent causes of HIV-associated deaths in the general population . This study, from a rural population, describes the causes of death in HIV-positive people and their HIV-negative controls . METHODS: A natural history cohort comprising HIV-1 infected participants and HIV-negative controls was established in rural Uganda in 1990 . Causes of death were determined by reviewing the premorbid clinical and laboratory findings and from information obtained from relatives . Blindness to the deceased's HIV serostatus was maintained throughout . RESULTS: In all, 78 deaths occurred over a 6-year period: 63 deaths occurred in the HIV-positive cases (53 prevalent and 10 incident cases) and 15 deaths in the HIV-negative controls . Of the prevalent cases, 56%, and 9% the incident cases enrolled died, compared with 7% of the HIV-negative controls . Of the 55 HIV-positive cases with sufficient data to establish cause of death, 52 (95%) were assessed as having HIV-associated deaths and 48 (87%) died in WHO stage 4 (AIDS) . The main causes of death were wasting syndrome (31%), chronic diarrhoea (22%), cryptococcal meningitis (13%) and chest infection (11%) . CONCLUSIONS: Our results represent an unbiased selection of deaths in a rural area . The HIV-positive cases have high death rates and die of HIV-related pathologies . The main causes of death reflect the WHO clinical case definition of AIDS . Cryptococcal meningitis is also a common cause of death in this populationPIP: A natural history cohort (NHC) of HIV-1-infected subjects and HIV-negative controls was established in rural Uganda in 1990 . By the end of 1996, 440 participants had enrolled in the cohort: 107 prevalent cases, 108 incident cases, and 225 HIV-negative controls . The authors report the causes of death among HIV-infected cohort members over the 6-year period ending December 1996 . Causes of death were determined by reviewing the premorbid clinical and laboratory findings, as well as from information obtained from relatives . All study clinic staff are blind to the HIV serostatus of participants in the NHC . 78 deaths occurred over the 6-year study period: 63 deaths among HIV-positive cases (53 prevalent and 10 incident cases) and 15 deaths among HIV-negative controls . 56% of prevalent cases, 9% of incident cases and 7% of controls died . Of the 55 HIV-positive cases with enough data to establish cause of death, 52 were determined to have HIV-associated deaths, of whom 48 died in World Health Organization stage 4 illness . Main causes of death were wasting syndrome (31%), chronic diarrhea (22%), cryptococcal meningitis (13%), and chest infection (11%) .

Antimicrob Agents Chemother, 1998 Oct, 42(10), 2630 - 2
In vitro and in vivo efficacy of the triazole TAK-187 against Cryptococcus neoformans; Schell WA et al.; Multiple isolates of Cryptococcus neoformans, including those with fluconazole resistance, were tested to assess the in vitro activity of the new triazole TAK-187 . MICs of TAK-187 were at least eightfold lower than those of fluconazole, and fungicidal concentrations for most isolates were 4 microg/ml or less . TAK-187 also was evaluated as intermittent therapy using two dosages in a rabbit model of experimental cryptococcal meningitis . Compared to daily treatment with fluconazole, as little as two doses of TAK-187 given 7 days apart were found to be effective . Plasma and cerebrospinal fluid TAK-187 concentrations were many times higher than MICs and fungicidal concentrations . Based upon its therapeutic efficacy and long half-life in the rabbit model, TAK-187 should be investigated for intermittent dosing in treatment or suppression of cryptococcal infections in humans.

Antimicrob Agents Chemother, 1998 Oct, 42(10), 2503 - 10
In vitro antifungal activities of a series of dication-substituted carbazoles, furans, and benzimidazoles; Del Poeta M et al.; Aromatic dicationic compounds possess antimicrobial activity against a wide range of eucaryotic pathogens, and in the present study an examination of the structures-functions of a series of compounds against fungi was performed . Sixty-seven dicationic molecules were screened for their inhibitory and fungicidal activities against Candida albicans and Cryptococcus neoformans . The MICs of a large number of compounds were comparable to those of the standard antifungal drugs amphotericin B and fluconazole . Unlike fluconazole, potent inhibitory compounds in this series were found to have excellent fungicidal activities . The MIC of one of the most potent compounds against C . albicans was 0.39 microg/ml, and it was the most potent compound against C . neoformans (MIC, </=0.09 microg/ml) . Selected compounds were also found to be active against Aspergillus fumigatus, Fusarium solani, Candida species other than C . albicans, and fluconazole-resistant strains of C . albicans and C . neoformans . Since some of these compounds have been safely given to animals, these classes of molecules have the potential to be developed as antifungal agents.

Antimicrob Agents Chemother, 1998 Oct, 42(10), 2495 - 502
Structure-in vitro activity relationships of pentamidine analogues and dication-substituted bis-benzimidazoles as new antifungal agents; Del Poeta M et al.; Twenty analogues of pentamidine, 7 primary metabolites of pentamidine, and 30 dicationic substituted bis-benzimidazoles were screened for their inhibitory and fungicidal activities against Candida albicans and Cryptococcus neoformans . A majority of the compounds had MICs at which 80% of the strains were inhibited (MIC80s) comparable to those of amphotericin B and fluconazole . Unlike fluconazole, many of these compounds were found to have potent fungicidal activity . The most potent compound against C . albicans had an MIC80 of </=0.09 microg/ml, and the most potent compound against C . neoformans had an MIC80 of 0.19 microg/ml . Selected compounds were also found to be active against Aspergillus fumigatus, Fusarium solani, Candida species other than C . albicans, and fluconazole-resistant strains of C . albicans and C . neoformans . It is clear from the data presented here that further studies on the structure-activity relationships, mechanisms of action and toxicities, and in vivo efficacies of these compounds are warranted to determine their clinical potential.

Indian Heart J, 1998 May-Jun, 50(3), 321 - 5
Pathology of the heart in acquired immunodeficiency syndrome; Lanjewar DN et al.; The spectrum of cardiac lesions in patients with acquired immunodeficiency syndrome in India is not described . To determine the extent of involvement of the heart with this disease, an autopsy study of 52 subjects having acquired immunodeficiency syndrome was carried out . Multiple sections were obtained from different anatomical parts of each heart . Forty-eight of the 52 hearts showed subtle microscopic changes, the most common being myocardial atrophy (48 cases), lymphocytic pericarditis (38 cases), fibrinous pericarditis (1 case), pericardial fibrosis (1 case), lymphocytic myocarditis (29 cases) and myocardial fibrosis (7 cases) . Cryptococcosis of the heart was noticed in two cases, while in one case toxoplasmic myocarditis was identified . In only one case clinical presentation of cardiac involvement (pericardial effusion) was noted, which indicates that in spite of the presence of significant pathology in the heart, overt cardiac manifestations are infrequently seen in patients with acquired immunodeficiency syndrome.

Infect Immun, 1998 Oct, 66(10), 5027 - 30
Organ-dependent variation of capsule thickness in Cryptococcus neoformans during experimental murine infection; Rivera J et al.; In studies of murine infection, the capsule thickness of Cryptococcus neoformans varied depending on the organ . The relative order of thickness was as follows: lung > brain > in vitro isolates . The differences in capsule thickness suggest that there are organ-related differences in the expression of genes responsible for capsule thickness.

Infect Immun, 1998 Oct, 66(10), 4994 - 5000
Interleukin-12 is essential for a protective Th1 response in mice infected with Cryptococcus neoformans; Decken K et al.; To analyze the roles of interleukin-12 (IL-12) and the IL-12-dependent Th1 response in resistance to Cryptococcus neoformans, we have established a chronic infection model in wild-type mice and in mice with targeted disruptions of the genes for the IL-12p35 and IL-12p40 subunits (IL-12p35(-/-) and IL-12p40(-/-) mice, respectively) as well as in mice with a targeted disruption of the IL-4 gene . Long-term application of exogenous IL-12 prevented death of infected wild-type mice for the entire period of the experiment (up to 180 days) but did not resolve the infection . Infected IL-12p35(-/-) and IL-12p40(-/-) mice died significantly earlier than infected wild-type mice, whereas infection of IL-4-deficient mice led to prolonged survival . Interestingly, infected IL-12p40(-/-) mice died earlier and developed higher organ burdens than IL-12p35(-/-) mice, which, for the first time in an infection model, suggests a protective role of the IL-12p40 subunit independent of the IL-12 heterodimer . The fungal organ burdens of IL-4-deficient mice and IL-12-treated wild-type mice were significantly reduced compared to those of untreated wild-type mice and IL-12-deficient mice . Histopathological analysis revealed reduction of the number of granulomatous lesions following treatment with IL-12 . Susceptibility of both IL-12p35(-/-) and IL-12p40(-/-) mice was associated with marginal production of gamma interferon and elevated levels of IL-4 from CD4(+) T cells, which indicates Th2 polarization in the absence of IL-12, whereas wild-type mice developed a Th1 response . Taken together, our data emphasize the essential role of IL-12 for protective Th1 responses against C . neoformans.

Rinsho Shinkeigaku, 1998 Apr, 38(4), 314 - 8
{Chronic Cryptococcal meningitis with CSF oligoclonal IgG band in a patient with Claude syndrome}; Kawanishi R et al.; We described a 61-year-old man who was diagnosed as having chronic cryptococcal meningitis, while he was hospitalized with Claude syndrome . The patient was admitted because of acute onset of gait disturbance . He had a tendency to fall down to his left side since he awoke in the morning of August 12, 1995 . On admission, he was mentally alert, showing a right oculomotor nerve palsy, gaze-evoked horizontal nystagmus in the left eye on the left lateral gaze, and incoordination of the left upper and lower extremities . In addition, he fell to the left side on standing up with feet together and with eyes closed . He had mild wild-based gait with a tendency to fall down to the left on tandem gait . Babinski sign was present on the left side . He did not have fever, nor meningeal signs, nor sensory abnormalities . X-ray films of the chest showed multiple nodular shadows consistent with pneumoconiosis . Cranial X-ray computed tomography and magnetic resonance imaging revealed a small lesion in the paramedian area of the midbrain on the right, consistent with an infarct . Cerebral arteriography revealed a stenosis in the proximal portion of the right posterior cerebral artery . Cerebrospinal fluid (CSF) showed a moderate mononuclear cell predominant pleocytosis, a moderate elevation of total protein, slightly reduced glucose values . Although the culture and India ink preparation of CSF were negative for cryptococcus in repeated studies, its antigen was positive both in the serum and CSF . In addition, the CSF showed an oligoclonal IgG band which was predominantly K type . After the antigen of Cryptococcus neoformans was added to the CSF in vitro, the oligoclonal IgG band was absorbed completely . The patient was treated with fluconazole (FLCZ), which did not cause any improvement of the CSF abnormalities, so that FLCZ was replaced by 5-flucytosine (5-FC) . Since the CSF abnormalities moderately improved with 5-FC, he was discharged on December 21, 1995 . After the 5-FC was discontinued, the CSF results slowly worsened over several months without any signs and symptoms of meningitis . He was hospitalized again on October 28, 1996 for treatment with both 5-FC and amphotericin B . Although the CSF abnormalities improved markedly, the meningitis was not cured . After he was discharged on February 1, 1997, he was treated with both 5-FC and FLCZ . Although his CSF abnormalities worsened mildly, he remained afebrile without meningeal signs and symptoms and led an ordinary life . In our patient it remained undetermined whether the Claude syndrome was caused by arteriosclerotic infarction, or vasculitis due to cryptococcal meningitis, or both . Asymptomatic chronic cryptococcal meningitis as observed in our patients is unusual . In addition, this is the second case after Porter et al (1977) that the oligoclonal IgG band in CSF proved to be related to cryptococcal infection.

Kyobu Geka, 1998 Aug, 51(9), 801 - 5
{Three resected cases of pulmonary cryptococcosis}; Fukuhara T et al.; Case 1 was a 53-year-old female who had a small nodule in the right S3 segment on chest CT . As she was not diagnosed by transbronchial lung biopsy (TBLB), open thoracotomy was performed . Case 2 was a 65-year-old female who had a nodule with pleural indentation in the right S6 segment . As this nodule showed difficulty to differentiate from small lung carcinoma, thoracoscopic surgery was performed . Case 3 was a 63-year-old female who had multiple lesions with cavity in the left S4 and S5 segments, which was preoperatively diagnosed by TBLB . She was performed thoracoscopic partial resection of the lingular segment because of poor response to antimycotic agents . All cases received preventive antimycotic agents for one or two months after the operation . There was no recurrence or postoperative meningitis . Thoracoscopic surgery is the effective procedure for the diagnosis and treatment of the localized pulmonary cryptococcosis.

Southeast Asian J Trop Med Public Health, 1998 Mar, 29(1), 105 - 7
Massive pulmonary cryptococcosis in an immunocompetent patient; Silachamroon U et al.; A 64-year-old man presented with progressive dyspnea . The symptom of severe hypoxia requiring mechanical ventilator, and bilateral pulmonary infiltrates on the chest film led to the clinical diagnosis of adult respiratory distress syndrome . Autopsy demonstrated widespread cryptococci and mucinous material in alveoli with mild inflammatory response.

J Antimicrob Chemother, 1998 Aug, 42(2), 217 - 20
In-vitro susceptibility of Cryptococcus neoformans isolates to fluconazole and itraconazole; Davey KG et al.; The in-vitro susceptibilities of 143 isolates of Cryptococcus neoformans, collected from British patients between 1994 and 1996, to fluconazole and itraconazole were compared with those of 36 isolates collected between 1971 and 1985, 41 isolates collected between 1986 and 1989, and 43 Ugandan isolates collected in 1996 . Testing was done with a broth microdilution method in YNB medium supplemented with glucose, incubation at 30 degrees C for 72 h, and an endpoint of 50% inhibition . The results showed that the MIC ranges, MIC50s and MIC90s of fluconazole and itraconazole for C . neoformans isolates from the UK have remained unchanged despite the recent widespread use of triazoles for long-term maintenance of patients with AIDS-associated cryptococcal meningitis . The MIC ranges, MIC50s and MIC90s of the 43 isolates from untreated Ugandan patients with AIDS were similar to those of the British isolates . Examination of our records for 1994-96 revealed six cases in which a four-fold or greater rise in the MIC of fluconazole was associated with relapsed cryptococcal meningitis.

J Clin Microbiol, 1998 Oct, 36(10), 3088 - 9
Utility of routine testing of bronchoalveolar lavage fluid for cryptococcal antigen; Kralovic SM et al.; All cryptococcal antigen (CrAg) testing performed at our institution between 1989 and 1994 was reviewed for utility of routinely testing of bronchoalveolar lavage fluid (BAL) for this antigen . Forty-two of 1,506 BAL specimens were positive . Seventeen of these were felt to represent false positives (sensitivity, 71%; positive predictive value, 0.59) . The data on CrAg in cerebrospinal fluid and serum and the fungal culture and histological results of BAL specimens did not support continued, routine testing of BALs for CrAg to diagnose cryptococcosis.

J Clin Microbiol, 1998 Oct, 36(10), 2950 - 6
Comparison of In vitro activities of the new triazole SCH56592 and the echinocandins MK-0991 (L-743,872) and LY303366 against opportunistic filamentous and dimorphic fungi and yeasts; Espinel-Ingroff A; The in vitro antifungal activities of SCH56592, MK-0991, and LY303366 against 83 isolates of Acremonium strictum, Aspergillus flavus, Aspergillus fumigatus, Aspergillus terreus, Bipolaris spp., Blastomyces dermatitidis, Cladophialophora bantiana, Fusarium oxysporum, Fusarium solani, Histoplasma capsulatum, Phialophora spp., Pseudallescheria boydii, Rhizopus arrhizus, Scedosporium prolificans, and Sporothrix schenckii were compared . The in vitro activities of these agents against 104 isolates of yeast pathogens of Candida spp., Cryptococcus neoformans, and Trichosporon beigelii were also compared . MICs were determined by following a procedure under evaluation by the National Committee for Clinical Laboratory Standards (NCCLS) for broth microdilution testing of the filamentous fungi (visual MICs) and the NCCLS M27-A broth microdilution method for yeasts (both visual and turbidimetric MICs) . The in vitro fungicidal activity of SCH56592 was superior (minimum fungicidal concentrations {MFCs}, 0.25 to 4 microgram/ml for 7 of 18 species tested) to those of MK-0991 and LY303366 (MFCs, 8 to >16 microgram/ml for all species tested) for the molds tested, but the echinocandins had a broader spectrum of fungicidal activity (MFCs at which 90% of strains are inhibited {MFC90s}, 0.5 to 4 microgram/ml for 6 of 9 species tested) than SCH56592 (MFC90s, 0.25 to 8 microgram/ml for 4 of 9 species tested) against most of the yeasts tested . Neither echinocandin had in vitro activity (MICs, >16 microgram/ml) against C . neoformans and T . beigelii, while the SCH56592 MICs ranged from 0.12 to 1.0 microgram/ml for these two species . The MICs of the three agents for the other species ranged from <0.03 to 4 microgram/ml . These results suggest that these new agents have broad-spectrum activities in vitro; their effectiveness in the treatment of human mycoses is to be determined.

J Clin Microbiol, 1998 Oct, 36(10), 2874 - 6
Fluconazole susceptibility testing of Cryptococcus neoformans: comparison of two broth microdilution methods and clinical correlates among isolates from Ugandan AIDS patients; Jessup CJ et al.; We compared the yeast nitrogen base (YNB) broth microdilution method with the National Committee for Clinical Laboratory Standards (NCCLS) M27-A microdilution reference method for measuring the in vitro susceptibility of Cryptococcus neoformans isolates to fluconazole . A total of 149 isolates of C . neoformans var . neoformans from Ugandan AIDS patients was tested by both methods . An overall agreement of 88% between the two microdilution methods was observed . All isolates grew well in both RPMI 1640 and YNB media, and MICs could be read after 48 h of incubation by both methods . The range of fluconazole MICs obtained with the YNB method was broader than that obtained with the NCCLS method . The extended range of MICs provided by the YNB method may be of clinical value, as it appears that the clinical outcome may be better among patients infected with strains inhibited by lower concentrations of fluconazole as determined by the YNB method . The YNB method appears to be a viable option for testing C . neoformans against fluconazole.

J Clin Microbiol, 1998 Oct, 36(10), 2817 - 22
Detection of resistance to amphotericin B among Cryptococcus neoformans clinical isolates: performances of three different media assessed by using E-test and National Committee for Clinical Laboratory Standards M27-A methodologies; Lozano-Chiu M et al.; Although reliable detection of resistance in vitro is critical to the overall performance of any susceptibility testing method, the recently released National Committee for Clinical Laboratory Standards M27-A methodology for susceptibility testing of yeasts discriminates poorly between resistant and susceptible isolates of Candida spp . We have previously shown that both substitution of antibiotic medium 3 for RPMI 1640 medium in the microdilution variant of the M27-A method and use of the E-test agar diffusion methodology permit detection of amphotericin B-resistant Candida isolates . To determine the relevance of these observations to Cryptococcus neoformans, we have evaluated the performances of both the M27-A and the E-test methodologies with this yeast using three different media (RPMI 1640 medium, antibiotic medium 3, and yeast nitrogen base) . As with Candida, we found that only antibiotic medium 3 permitted consistent detection of resistant isolates when testing was performed in broth by the M27-A method . When testing was performed by the E-test agar diffusion method, both RPMI 1640 medium and antibiotic medium 3 agar permitted ready detection of the resistant isolates . Reading of the results after 48 h of incubation was required for testing in broth by the M27-A method, while the MIC could be determined after either 48 or 72 h when the agar diffusion method was used.

Antimicrob Agents Chemother, 1998 Sep, 42(9), 2274 - 8
Sordarins: A new class of antifungals with selective inhibition of the protein synthesis elongation cycle in yeasts; Dominguez JM et al.; GR135402, a sordarin derivative, was isolated in an antifungal screening program . GR135402, sordarin, and derivatives of both compounds were evaluated for their ability to inhibit cell-free translational systems from five different pathogenic fungi (Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, and Cryptococcus neoformans) . The activity profile of GR135402 is extended to other chemical compounds derived from sordarin . Experimental results indicate that sordarin analogs exert their antifungal effects by specifically inhibiting the protein synthesis elongation cycle in yeasts but do not affect protein synthesis machinery in mammalian systems . Intrinsically resistant strains owe their resistance to differences in the molecular target of sordarins in these strains . Preliminary studies performed to elucidate the mode of action of this new class of antifungal agents have shown that the putative target of sordarins is one of the protein synthesis elongation factors.

Am J Gastroenterol, 1998 Sep, 93(9), 1592 - 3
Primary intestinal cryptococcosis mimicking adenomatous polyp in an HIV-negative patient; Melato M et al.; Primary cryptococcal infection is thought to arise in the lungs, whereas secondary lesions may be found anywhere in the body . Because intestinal involvement is rare, especially in nonimmunocompromised patients, little is known about this localization . Nevertheless, the intestinal tract has long been suggested a possible portal of entry of Cryptococcus neoformans, although the hypothesis has never been sufficiently documented . We report an isolated cryptococcosis of the sigmoid colon mimicking an adenomatous polyp . The lesion has an endoscopic interest, being the first of its kind reported in the literature, and a more important pathogenic interest, as it highlights a further pathway of cryptococcal infection, one of major importance in immunocompromised patients.

J Infect Dis, 1998 Sep, 178(3), 803 - 14
Interleukin-15 induces antimicrobial activity after release by Cryptococcus neoformans-stimulated monocytes; Mody CH et al.; A newly described cytokine, interleukin (IL)-15, shares many activities with IL-2; however, little is known about the stimuli for release of IL-15, and its role in antimicrobial host defense has not previously been demonstrated . This study found that Cryptococcus neoformans is a potent stimulus for the release of biologically active IL-15 from monocytes . Both IL-15 and IL-2 made significant contributions to lymphocyte proliferation and lymphocyte-mediated anticryptococcal activity to encapsulated and acapsular C . neoformans . IL-15 restored lymphocyte proliferation and anticryptococcal activity that had been abrogated by blocking IL-2 . IL-15 also enhanced the anticryptococcal activity of lymphocytes but did not enhance the activity of monocytes . This suggests that IL-15 and IL-2 cooperate for lymphocyte activation and proliferation in vitro and demonstrates that IL-15 can induce antimicrobial activity . Taken together, these data suggest that microbes, and in particular C . neoformans, are an important stimulus for IL-15 and that IL-15 may have an important role in induction of antimicrobial effector mechanisms.

AIDS, 1998 Aug 20, 12(12), 1491 - 4
HIV combination therapy: partial immune restitution unmasking latent cryptococcal infection; Woods ML 2nd et al.; OBJECTIVE: To describe two cases of cryptococcal meningitis and one re-exacerbation of Cryptococcus-associated meningitis occurring in temporal association with commencement of highly active antiretroviral therapy (HAART) in patients with advanced HIV infection (CD4 cells < 50 x 10(6)/l), which suggests that partial immune restitution can facilitate development of clinically apparent meningitis in response to Cryptococcus or its antigen . DESIGN: All HIV-infected patients with culture-proven cryptococcal meningitis diagnosed at a tertiary referral centre specialist infectious diseases unit from 1 January 1996 to 31 December 1996 were reviewed to examine the clinical and immunological parameters prior to and after commencing antiretroviral therapy . RESULTS: Three patients were diagnosed with clinically apparent meningitis within 7-39 days of changing or altering antiretroviral combination therapy consisting of zidovudine or stavudine, in combination with lamivudine and saquinavir . All patients had CD4 cell counts below 50 x 10(6)/l at initiation of therapy . Following institution of HAART, evidence of immune restitution was suggested by the following: (i) significant increases (3.7-14-fold) in numbers of CD4 cells (all three patients), (ii) significantly reduced (> 2-4 log10 reduction) HIV viral loads (two out of three patients), and (iii) prominent inflammatory changes in cerebrospinal fluid (white blood cells > 10 x 10(6)/l) at diagnosis (two out of three patients) . CONCLUSIONS: Our report suggests that in patients with advanced HIV infection, partial immune restitution induced by HAART can precipitate onset of clinically apparent meningitis in those patients with latent cryptococcal central nervous system infection or with residual cryptococcal antigen present in the cerebrospinal fluid.

J Pharm Sci, 1998 Sep, 87(9), 1144 - 8
Liposomes, a potential immunoadjuvant and carrier for a cryptococcal vaccine; Lambros MP et al.; Mice immunized with a cryptococcal culture filtrate antigen (CneF) emulsified in complete Freund's adjuvant (CFA) develop an anticryptococcal cell-mediated immune response (CMI) . CMI is detected by delayed-type hypersensitivity (DTH) reactions and by enhanced clearance of Cryptococcus neoformans from infected tissues . The objective of this research was to evaluate anticryptococcal DTH reactivity and clearance of cryptococci from groups of mice immunized with CneF encapsulated into liposomes (CneF-liposome) and compare the results to results from mice immunized with CneF-CFA . CBA/J mice were injected subcutaneously with vaccines or control formulations (saline-liposome or saline-CFA) . Six days later the mice were footpad tested to assess their DTH response to CneF or the animals were challenged intravenously with 10(5) viable C . neoformans to determine clearance of infection . Clearance was evaluated 7 days later by enumeration of cryptococcal colony forming units (CFU) in lungs, spleens, livers, and brains of the infected mice . The CneF-liposome formulation induced a positive anticryptococcal DTH response and elicited increased clearance of C . neoformans from tissues as compared to mice treated with saline-liposome . Even though the CneF-liposome preparation did not induce as strong of a DTH response or as much protection as did CneF-CFA, our results indicate that liposomes are promising carriers for immunization with cryptococcal antigen and that such immunization can provide some protection to subsequent infection with C . neoformans.

Rev Med Chil, 1991 Nov, 119(11), 1291 - 5
{A patient with AIDS: infection associated with pneumocystis carinii and cryptococcus neoformans}; Fica A et al.; A patient with AIDS developed pulmonary infiltrates typical of infection by P . carinii . However, therapy for this agent was unsuccessful . Further studies including blood cultures revealed the presence of C neoformans . Specific therapy for this agent led to recovery . Concomitant opportunistic infections may cause diagnostic and therapeutic problems in patients with AIDS.

Mycoses, 1998, 41 Suppl 1, 86 - 9
{Epidemiology of aspergillosis and cryptococcosis in Germany}; Polak A et al.; The results of an enquiry concerning aspergillosis and cryptococcosis for the period 1992-1997 in Germany are reported and analysed.

J Appl Microbiol, 1998 Jun, 84(6), 937 - 44
Ecomycins, unique antimycotics from Pseudomonas viridiflava; Miller CM et al.; A novel family of peptide antimycotics, termed ecomycins, is described from Pseudomonas viridiflava, a plant-associated bacterium . Ecomycins B and C have molecular masses of 1153 and 1181 . They contain equimolar amounts of a beta hydroxyaspartic acid, homoserine, threonine, serine, alanine, glycine and one unknown amino acid . Fatty acids were detectable after hydrolysis, methylation and gas chromatography and mass spectroscopy . The ecomycins have significant bioactivities against a wide range of human and plant pathogenic fungi . The minimum inhibitory concentration values for ecomycin B were 4.0 micrograms ml-1 against Cryptococcus neoformans and 31 micrograms ml-1 against Candida albicans . Pseudomonas viridiflava also produces what appears to be syringotoxin, an antifungal lipopeptide previously described from Ps . syringae.

Mycoses, 1998 May-Jun, 41(5-6), 199 - 202
Enhanced activity of antifungal drugs by lysozyme against Cryptococcus neoformans; Nakamura Y et al.; The in vitro susceptibility of 16 isolates of Cryptococcus neoformans to three antifungal drugs and lysozyme in combination was determined using an urea broth microdilution method . The antifungal activities of each drug alone against 16 isolates of Cr . neoformans were determined as mean minimal inhibitory concentrations (MICs) . MICs of fluconazole, itraconazole and terbinafine were 2.0 micrograms ml-1, 0.004 microgram ml-1 and 0.25 microgram ml-1, respectively . Lysozyme alone inhibited the growth of Cr . neoformans in a dose-dependent manner, although the lysozyme was unable to kill the cells of Cr . neoformans at the highest concentration of 20 micrograms ml-1 . The mean MICs of fluconazole, itraconazole and terbinafine in combination with lysozyme were 0.13 microgram ml-1, 0.004 microgram ml-1 and 0.03 microgram ml-1 respectively . The antifungal activity of fluconazole and terbinafine in combination with lysozyme against Cr . neoformans was greatly enhanced compared with that of each drug alone . Itraconazole was unable to enhance the antifungal activity, as it demonstrated higher activity against Cr . neoformans when alone rather than in combination . Lysozyme was confirmed to enhance the antifungal activity of fluconazole and terbinafine in vitro.

Mycoses, 1998 May-Jun, 41(5-6), 195 - 8
Rapid and easy method to extract and preserve DNA from Cryptococcus neoformans and other pathogenic yeasts; Sansinforiano ME et al.; The mucopolysaccharide capsule of Cryptococcus neoformans and other pathogenic yeasts prevent the extraction of DNA from these important zoonotic agents . We report that the use of a lysis buffer containing a high concentration of urea is an easy, efficient and time-saving technique to obtain high yields of good-quality DNA for molecular diagnosis . The use of urea also prevents the degradation of DNA during storage of samples at room temperature for up to 6 months.

Arch Pharm (Weinheim), 1998 Jun, 331(6), 225 - 7
Allylamine type xanthone antimycotics; Salmoiraghi I et al.; A number of xanthone derivatives bearing the basic chain of naftifine and butenafine antimycotics in 1, 2, 3, and 4 nuclear positions are described . The in vitro antifungal activity against representative strains of molds and yeasts is reported . Only butenafine xanthone analogues show significant activity against Cryptococcus neoformans, in particular the regioisomer 4d (1.5 micrograms/ml).

Infect Immun, 1998 Sep, 66(9), 4324 - 30
Involvement of C3a and C5a in interleukin-8 secretion by human polymorphonuclear cells in response to capsular material of Cryptococcus neoformans; Vecchiarelli A et al.; In a previous paper we demonstrated that human polymorphonuclear cells (PMN) in the presence of normal human serum (NHS) secrete proinflammatory cytokines in response to Cryptococcus neoformans or its major capsular component, glucuronoxylomannan (GXM) . The hypothesis that activation of the complement system could be responsible for the observed phenomenon is supported by the fact that encapsulated and acapsular C . neoformans isolates are activators of the complement system and, in particular, large encapsulated isolates are powerful activators . In the present study we demonstrate that (i) interleukin-8 (IL-8) release in response to acapsular or encapsulated strains of C . neoformans is significantly reduced in the presence of heat-inactivated serum rather than NHS and is completely abrogated in the absence of human serum; (ii) GXM-induced IL-8 release is strictly dependent on the presence of NHS, is inhibited by specific antibodies to either C3a and C5 complement components, and is completely abrogated by the combined use of these antibodies; (iii) the addition of purified C3a and C5a directly stimulates IL-8 release by PMN; and (iv) monoclonal antibody to GXM in combination with GXM or encapsulated C . neoformans potentiates IL-8 release by PMN . These data shed light on the mechanism involved in GXM-induced IL-8 secretion by PMN, provide an additional potential role for complement in the control of C . neoformans infections, and suggest a complex interplay between the complement system, humoral immunity, and cytokine regulation.

Infect Immun, 1998 Sep, 66(9), 4169 - 75
Ferrous iron uptake in Cryptococcus neoformans; Jacobson ES et al.; Previous studies have implicated ferric reduction in the iron uptake pathway of the opportunistic pathogen Cryptococcus neoformans . Here we studied iron uptake directly, using 55Fe in the presence of reductants . Uptake was linear with respect to time and number of yeast cells . The plot of uptake versus concentration exhibited a steep rise up to about 1 microM, a plateau between 1 and 25 microM, and a second steep rise above 25 microM, consistent with high- and low-affinity uptake systems . A Km for high-affinity uptake was estimated to be 0.6 microM Fe(II); 1 microM was used for standardized uptake assays . At this concentration, the uptake rate was 110 +/- 3 pmol/10(6) cells/h . Iron repletion (15 microM) and copper starvation drastically decreased high-affinity iron uptake . Incubation at 0 degreesC or in the presence of 2 mM KCN abolished high-affinity iron uptake, suggesting that uptake requires metabolic energy . When exogenous reducing agents were not supplied and the culture was washed free of secreted reductants, uptake was reduced by 46%; the remaining uptake activity presumably was dependent upon the cell membrane ferric reductase . Further decreases in free Fe(II) levels achieved by trapping with bathophenanthroline disulfonate or reoxidizing with potassium nitrosodisulfonate reduced iron uptake very drastically, suggesting that it is the Fe(II) species which is transported by the high-affinity transporter . The uptake of Fe was stimulated two- to threefold by deferoxamine, but this increment could be abolished by copper starvation or inhibition of the ferric reductase by Pt, indicating that Fe solubilized by this molecule also entered the reductive iron uptake pathway.

Infect Immun, 1998 Sep, 66(9), 4018 - 24
Decreased resistance to primary intravenous Cryptococcus neoformans infection in aged mice despite adequate resistance to intravenous rechallenge; Aguirre KM et al.; It is often stated that impaired immune functions in the aged underlie their greater susceptibility to infections . Indeed, in many experimental settings, T-cell responses in aged mice have been shown to be deficient compared with those from young adults . Nonetheless, there are very few examples where a greater susceptibility to infection in aged mice has been demonstrated to result from impaired T-cell function . The clinical importance of understanding the basis for increased susceptibility to infection that accompanies advanced age dictates a need for experimental models with which to study the effect that aging has on immunological resistance to infection . This study was undertaken to investigate whether aged mice were less resistant than young adult control mice to infection with the fungus Cryptococcus neoformans . After a primary intravenous challenge with yeast, aged mice died sooner and developed higher organ burdens of yeast than did young adults . Deficient in vitro responses were observed in T cells from aged mice; however, greater susceptibility to intravenous infection appeared not to result from less effective T-cell-dependent resistance in vivo . In fact, T-cell-replete aged mice were more susceptible to intravenous cryptococcal infection than were T-cell-depleted young adults . Furthermore, aged mice were as resistant to primary pulmonary challenge with Cryptococcus as were young adults . Similarly, vaccinated aged mice were as resistant to rechallenge as were young adult counterparts . Therefore, despite demonstrably deficient in vitro responses of T cells from aged mice, their T-cell-dependent resistance to C . neoformans is as effective as that of young adults.

J Immunol, 1998 Aug 15, 161(4), 1829 - 36
Aspects of antigen mimicry revealed by immunization with a peptide mimetic of Cryptococcus neoformans polysaccharide; Valadon P et al.; We have recently identified peptide mimetics of the Cryptococcus neoformans capsular polysaccharide by screening phage display peptide libraries . 2H1, one of a large family of mAbs against the glucuronoxylomannan fraction (GXM), is highly protective and binds several peptide motifs . This study analyzes the immunologic properties of P601E (SYSWMYE), a peptide from the low affinity motif (W/YXWM/LYE) that has an extended cross-reactivity among anti-GXM mAbs and whose binding correlates with the protective potential of mAbs in experimental infection . P601E is a mimetic, since it competes for GXM binding to 2H1, but not a mimotope, since it does not elicit an anti-GXM response . Sequence analysis of 14 anti-P601E mAbs indicates that anti-P601E mAbs elicited in BALB/c mice have an order of homology with 2H1 of V kappa > J kappa >> V(H) > J(H) > D . Further screening of a peptide library with anti-P601E mAbs isolated peptides having a motif almost identical to the peptide motif selected by 2H1 . When these results are compared to the crystal structure of a related peptide in complex with 2H1, there is a clear correlation between the ability to elicit V region components of 2H1 Ab and peptide association with the V region, suggesting that the completeness of the fit in the binding site is an important driving force for mimicry . As a consequence, improving affinity of a mimetic for the Ab binding site seems to be the most logical way to insure that all of the appropriate V region segments are elicited and that useful mimotopes are created.

J Clin Invest, 1998 Aug 15, 102(4), 663 - 70
Granulocyte colony-stimulating factor administration to HIV-infected subjects augments reduced leukotriene synthesis and anticryptococcal activity in neutrophils; Coffey MJ et al.; Neutrophil (PMN) dysfunction occurs in HIV infection . Leukotrienes (LT) are mediators derived from the 5-lipoxygenase (5-LO) pathway that play a role in host defense and are synthesized by PMN . We investigated the synthesis of LT by PMN from HIV-infected subjects . There was a reduction (4.0+/-1.3% of control) in LT synthesis in PMN from HIV-infected compared with normal subjects . This was associated with reduced expression of 5-LO-activating protein (31.2+/-9.6% of normal), but not of 5-LO itself . Since HIV does not directly infect PMN, we considered that these effects were due to reduced release of cytokines, such as granulocyte colony-stimulating factor (G-CSF) . We examined the effect of G-CSF treatment (300 microgram daily for 5 d) on eight HIV-infected subjects . PMN were studied in vitro before therapy (day 1) and on days 4 and 7 . LTB4 synthesis was increased on day 4 of G-CSF treatment, and returned toward day 1 levels on day 7 . 5-LO and 5-LO-activating protein expression were increased in parallel . As a functional correlate to this increase in PMN LT synthesis by G-CSF, we examined the effects on killing of Cryptococcus neoformans . Anticryptococcal activity of PMN from HIV-infected subjects was less than that of PMN from normal subjects . G-CSF treatment improved fungistatic activity of PMN . This increase in antifungal activity was attenuated by in vitro treatment with the LT synthesis inhibitor, MK-886 . In conclusion, PMN from HIV-infected subjects demonstrate reduced 5-LO metabolism and antifungal activity in vitro, which was reversed by in vivo G-CSF therapy.

Clin Infect Dis, 1998 Aug, 27(2), 260 - 4
Flucytosine monotherapy for cryptococcosis; Hospenthal DR et al.; Flucytosine (5-FC) monotherapy for cryptococcosis is not advocated because drug resistance emerges during therapy . Reported documentation of this widely accepted belief is surprisingly scarce . Therefore, we reviewed our experience with 5-FC monotherapy for 27 patients treated between 1968 and 1973 . Patients were selected on the basis of criteria associated with good prognosis . In this group, 5-FC monotherapy resulted in cure in eight cases and improvement in two . Overall, response was seen in 10 (43%) of 23 evaluable patients . Therapy failed for 13 patients, including 5 who relapsed, 2 who had partial responses, and 6 without response . Resistance was noted to have developed in isolates from six (50%) of 12 patients for whom therapy failed . Although the 57% failure rate associated with 5-FC alone precludes its use as monotherapy, our study did show that this treatment was well tolerated and that failure was not invariably associated with development of resistance.

Int J Food Microbiol, 1998 Jun 16, 41(3), 185 - 94
Yeast profile in Gouda cheese during processing and ripening; Welthagen JJ et al.; The yeasts present in Gouda cheese during processing were monitored in a single cheese factory during a 32 day ripening period . Despite the predominance of lactic acid bacteria during Gouda making, yeasts played a significant role in the ripening process in reaching counts as high as 10(5) cfu g(-1) at the later stages of ripening . The increase in the number of yeasts corresponded with the depletion in lactose content and the simultaneous stabilization of lactic acid bacteria numbers . The sources of yeast contamination which may lead to contamination of the curd were also determined . The brine and equipment surfaces were responsible for the highest yield of contaminating yeasts . A diverse variety of 23 yeast species representing 13 genera were present in the factory environment, during processing and ripening . Samples were taken at critical control points in the manufacturing process and analysed after incubation at 25 degrees C for 96 h . Although a broad spectrum of yeasts were found in Gouda cheese, Debaryomyces hansenii was the most abundant yeast isolated . Other species encountered were Saccharomyces cerevisiae and Yarrowia lipolytica, Kluyveromyces marxianus, Torulaspora delbrueckii, Rhodotorula glutinis, Cryptococcus albidus and Candida catenulata.

J Clin Microbiol, 1998 Sep, 36(9), 2742 - 4
Identification of Trichosporon asahii by PCR based on sequences of the internal transcribed spacer regions; Sugita T et al.; Trichosporon asahii is a major causative agent of deep-seated trichosporonosis, which has a high mortality rate . To detect T . asahii, we have developed specific oligonucleotide primers based on the internal transcribed spacer regions of this organism's genome . Amplification products were selectively obtained from only T . asahii DNA; the DNAs of other Trichosporon species, as well as those of medically relevant yeasts such as Candida albicans, Cryptococcus neoformans, and Malassezia furfur, were not amplified . This detection system will be useful as a microbiological tool for the diagnosis of trichosporonosis.

J Am Acad Dermatol, 1998 Aug, 39(2 Pt 2), 334 - 7
Plaque-like erythema with milia: a noninfectious dermal mucinosis mimicking cryptococcal cellulitis in a renal transplant recipient; Carrington PR et al.; Cellulitis of the skin is most commonly a bacterial infection caused by either staphylococcal or streptococcal species . However, in immunocompromised patients, cellulitis may be a harbinger of more ominous disease . In these patients, rapid clinical diagnosis and treatment is necessary . We describe an immunosuppressed renal transplant recipient with the clinical diagnosis of cryptococcal cellulitis who was found to have a heretofore unreported dermal mucinosis of the upper chest associated with milia . We believe that cyclosporine was a significant pathogenic factor . This entity, which should be included in the clinical differential diagnosis of cryptococcal cellulitis, has a benign nature and does not require treatment.

J Clin Pharmacol, 1998 Jul, 38(7), 583 - 92
AmBisome (liposomal amphotericin B): a comparative review; Boswell GW et al.; AmBisome (NeXstarPharmaceuticals, San Dimas, CA) is a unilamellar liposomal formulation of amphotericin B that was recently approved for use as empirical treatment for presumed fungal infections in febrile neutropenic patients and for aspergillosis, candidiasis, and cryptococcosis infections refractory to amphotericin B . It is a small closed microscopic sphere (<100 nm in diameter) with an inner aqueous core (i.e., a true liposome) . AmBisome remains as an intact sphere in vitro and for prolonged periods of time in vivo during the processes of systemic transport and pharmacologic action . As a consequence of its size and in vivo stability, AmBisome has physiochemical properties and a pharmacokinetic profile that are considerably different from those of currently available lipid-complexed amphotericin B formulations, with greatly increased area under the plasma concentration-time curve and much lower clearance at equivalent doses . AmBisome liposomes can be seen to accumulate at sites of fungal infection . Disruption of AmBisome liposomes occurs after attachment to the fungal cell wall and results in amphotericin B binding to fungal cell membrane ergosterol with subsequent cell lysis . AmBisome has been shown to penetrate the cell wall of both extracellular and intracellular forms of susceptible fungi.

Nippon Ishinkin Gakkai Zasshi, 1998, 39(3), 123 - 7
{Three-dimensional reconstruction of mitotic cells of Cryptococcus neoformans based on serial section electron microscopy}; Mochizuki T; Ultra-thin serial sections of mitotic cells of Cryptococcus neoformans were observed by transmission electron microscope and computer-aided reconstruction of three-dimensional models were performed to understand the kinetics of the nucleus and mitochondria during mitosis . The separation of chromosomes occured in the protrusion of the nucleus extending in the bud (=daughter cell), followed by one set of chromosomes being moved back into the mother cell . These findings were similar to those reported in the heterobasidiomycetous yeasts . No giant mitochondrion, composed of a coalescence of all the mitochondria, was found . Volumetric analysis revealed the fluctuations in the mitochondrial volume: total cytoplasmic volume ratio were minimum during mitosis . In addition, methods, usefulness and limitations of observation of ultra-thin serial sections for understanding the morphology of cells and intracellular organelles were reviewed.

Ann Hematol, 1998 Jun, 76(6), 283 - 6
Cryptococcosis in Hodgkin's disease: description of two cases and review of the literature; Korfel A et al.; Systemic mycosis caused by Cryptococcus neoformans frequently becomes life threatening in patients with cellular immunodeficiencies . In contrast to AIDS patients, there are only a few reports of concurrent systemic cryptococcosis in patients with Hodgkin's disease (HD) . Only two of 75 (2.7%) patients with HD who were consecutively admitted to our hospital in the past decade developed Cryptococcus neoformans infection . Both had stage IVB (Ann Arbor) HD with bone marrow involvement and absolute lymphopenia (< 1/nl) . We have reviewed the literature and analyzed the data of 54 cases with concurrent cryptococcosis and HD . Presence of HD for > or = 12 months, stage IV disease, absolute lymphopenia (< 1/nl), and extensive pretreatment were the most common features among these patients and must be regarded as predisposing for acquiring a cryptococcal infection . In our patients antimycotic therapy was successful using liposomal amphotericin B (lipAmB) simultaneously with cytotoxic therapy for HD . Drug level measurements performed in one patient revealed a higher level of amphotericin B in CSF when the liposomal formulation was administered as compared with the level in CSF after administration of conventional amphotericin B . To our knowledge, this is the first report on antimycotic treatment of cryptococcosis with lipAmB in patients with HD . Regarding the favorable therapeutic index of lipAmB as compared with conventional amphotericin B, the drug should be considered as a less toxic and perhaps more effective alternative in the therapy of acute cryptococcosis, especially when cytotoxic treatment is administered simultaneously.

Ann Trop Paediatr, 1998 Mar, 18(1), 45 - 8
Idiopathic CD4+ T-lymphocytopenia in a child with disseminated cryptococcosis; Menon BS et al.; We describe a Malay girl with disseminated cryptococcosis affecting the lungs, liver, lymph nodes and bones . The diagnosis was made by culture of the bone marrow . Tests of immune function showed that she was HIV-negative but the CD4 percentage was persistently low . Idiopathic CD4+ T-lymphocytopenia was diagnosed . The child died despite two courses of anti-fungal therapy.

Mycopathologia, 1997-98, 140(3), 115 - 20
Co-infection by Cryptococcus neoformans and Mycobacterium avium intracellulare in AIDS . Clinical and epidemiological aspects; Arasteh K et al.; In the observation of various opportunistic pathogens in HIV-positive persons, co-infection by Cryptococcus neoformans together with Mycobacterium avium intracellulare was found if there was a CD4 lymphocyte count as low as 3-20/microliters . In 1540 HIV-positive patients under treatment at a Berlin hospital (Auguste-Viktoria-Krankenhaus) during 1985-1994, all AIDS-relevant diseases were examined in a multivariate analysis as variables of influence on the manifestation of a systemic Mycobacterium avium complex (MAC) infection . The analysis involved data on 36 cases of cryptococcosis and 202 cases with a typical clinical course in whom MAC had been detected at sterile body sites . As significant and independent factors of influence, the following were identified: C . neoformans infection, wasting syndrome, lower age, low CD4 lymphocyte count and preceding Pneumocystis carinii pneumonia (PcP) prophylaxis . Cryptococcosis ranged first with an ods ratio of 2.75 . The concomitant manifestation of cryptococcosis and systemic MAC infection in six patients is shown . Because both opportunists, C . neoformans and avian mycobacteria, may have their common habitat in droppings of defined species of pet birds, a common source of infection deserves further clinical and epidemiological attention.

Carbohydr Res, 1998 Jan, 306(1-2), 315 - 30
Structural characterization of the galactoxylomannan of Cryptococcus neoformans Cap67; Vaishnav VV et al.; The galactoxylomannan (GalXM) obtained from the culture supernatant of an acapsular mutant of Cryptococcus neoformans Cap67 was purified by Concanavalin A affinity, ion-exchange, and gel-filtration chromatographies . The structure of GalXM was determined by methylation analysis and by 1D and 2D NMR spectroscopic studies of the intact polysaccharide and of the oligosaccharide fragments generated by Smith degradation and by acetolysis . GalXM is a complex polysaccharide with an alpha-(1-->6) -galactan backbone . The polysaccharide is branched at c-3 of alternate Gal units of the backbone . C-3 is the point of attachment of the oligosaccharide side chains comprised of alpha-D-Man- (1-->3)-alpha-D-Man-(1-->4)- beta-D-Gal-substituted with zero to three terminal beta-Xyl residues as shown in the following structure: {formula: see text}.

Curr Genet, 1998 Jul, 34(1), 60 - 6
Construction of stable episomes in Cryptococcus neoformans; Varma A et al.; We report the generation of stable plasmids constructed by inserting specific DNA sequences into previously known unstable vectors . These sequences were obtained from a DNA library recovered from a previously reported stable minichromosome created by electroporative transformation in Cryptococcus neoformans (Varma and Kwon-Chung 1994) . A 6-kb insert from this minichromosome significantly enhanced both the frequencies at which URA5 transformants were obtained as well as the stability of their uracil prototrophy on non-selective media . A 1.5-kb sequence of this insert contained telomeric sequence repeats which when introduced into plasmids resulted in significant increases in transformation frequency . A 1081-bp sequence (STAB), present in the remainder of the insert, had an ARS-like function enhancing the episomal maintenance of plasmids in the transformants regardless of the gene (ADE2/URA5) used as a selection marker.

Intern Med, 1998 Jun, 37(6), 534 - 7
Cryptococcal pleural effusion in a patient with chronic renal failure receiving long-term corticosteroid therapy for rheumatoid arthritis; Fukuchi M et al.; A 52-year-old woman with a seven-year history of rheumatoid arthritis (RA) was transferred to our department with chronic renal failure to undergo hemodialysis . She had been treated with prednisolone for a long time, and had renal amyloidosis secondary to RA . During her hospitalization, a left pleural effusion developed . Pleural fluid cultured positive for Cryptococcus neoformans (CN), and the CN antigen was detected in both pleural fluid and serum . Chest computerized tomography revealed an infiltrate shadow in the left lower lung field suggestive of CN infection . This was successfully treated with anti-fungal agents . Pleural effusion is an unusual manifestation of pulmonary cryptococcosis . We should consider a diagnosis of CN infection when pleural effusion is observed in compromised patients such as those receiving a long-term corticosteroid treatment.

J Cutan Med Surg, 1998 Jul, 3(1), 43 - 5
Cutaneous cryptococcosis mimicking basal cell carcinoma in a patient with AIDS; Ingleton R et al.; BACKGROUND: Cryptococcosis is an opportunistic infection caused by the encapsulated yeast Cryptococcus neoformans . This ubiquitous organism has emerged as a frequent finding in immunosuppressed patients, especially those with underlying malignancies, organ transplants, and the acquired immune deficiency syndrome (AIDS) . Cutaneous manifestations of cryptococcosis occur in 10 to 15% of patients having systemic involvement . These skin lesions may simulate a variety of different disease entities . METHODS: A case of crytococcosis mimicking a basal cell carcinoma is the subject of a case report presentation . RESULTS: A case of cutaneous cryptococcosis mimicking basal cell carcinoma occurred in a patient with AIDS, who did not appear to have dissemination, but was treated aggressively to stem possible occult systemic disease . CONCLUSION: Cutaneous crytococcosis may mimic other dermatologic disorders.

J Med Assoc Thai, 1998 Jun, 81(6), 462 - 7
Primary cryptococcal infection of the larynx in a patient with AIDS: a case report; Chongkolwatana C et al.; Primary laryngeal cryptococcosis was reported in a 42-year-old man with AIDS . The patient also had pulmonary tuberculosis and hydropneumothorax as a complication . Serological tests and/or cultures from blood, CSF, urine and pleural fluid were all negative for cryptococcus . He was successfully treated with oral fluconazole for 8 weeks to clear the infection and remained clear in the follow-up period 9 months after treatment.

Allergol Immunopathol (Madr), 1998 May-Jun, 26(3), 144 - 50
{Prevention and treatment of opportunistic infections}; Nevot Falco S et al.; An opportunist infection (OI) is understood to be an infection produced by microorganisms that invade a host with impaired immune capacity, such as children with HIV infection . The adequate treatment and chemoprophylaxis of these infections has improved the prognosis of their evolution, although they still present a high morbidity and mortality when they occur . In this sense, the introduction of triple therapy (new antiretroviral inhibitors and protease inhibitors) is likely to produce a prompt decrease in the incidence of OI because of the regression in the degree of immunosuppression that it induces . The degree of immunosuppression is determined by the number of CD4 lymphocytes, the most reliable marker for assessment . Normal CD4 lymphocytes values are different for each age group and have important connotations for the prophylactic measures to be used at each moment depending on the CD4 lymphocyte count . Opportunist infections influence the quality of life of patients . More than 100 microorganisms, including bacteria, viruses, fungi and protozoa, cause OI . This paper describes primary and secondary prophylaxis as well as the treatment of the most frequent opportunist infections (Pneumocystis carinii pneumonia, bacterial infections, Cryptococcus neoformans, Cytomegalovirus, Herpes simple, Varicella-zoster virus . Toxoplasmosis, Mycobacterium tuberculosis, M . avium-intracellulare, M . kansasii).

Rev Inst Med Trop Sao Paulo, 1997 Nov-Dec, 39(6), 323 - 5
Cerebrospinal fluid profiles in acquired immunodeficiency syndrome with and without neurocryptococcosis; Garlipp CR et al.; Cryptococcosis is one of the most common fungal infections of the central nervous system (CNS) in AIDS patients and meningoencephalitis or meningitis is a frequently observed manifestation . However, systematic studies of cerebrospinal fluid (CSF) composition from AIDS patients with CNS cryptococcosis have been few . CSF samples from 114 HIV seropositive patients whose clinical complaint suggested CNS involvement, were analyzed; 32 samples from patients diagnosed as having neurocryptococcosis (Group 1) and 82 samples from patients with no identified neurological disfunction (Group 2) . Based on cytological and biochemical results, two distinct profiles were observed: Normal (Group 1 = 31%, Group 2 = 39%); Abnormal (Group 1 = 69%, Group 2 = 61%) . Lymphocytes were the most frequent cells in both groups . Our CSF cytological and biochemical findings showed that in AIDS patients liquoric abnormalities are quite frequent, non-specific and difficult to interpret . In these circumstances a systematic search to identify the etiologic agent using microbiological and/or immunological assays must be routinely performed.

Am J Med Sci, 1998 Jul, 316(1), 60 - 4
Disseminated cryptococcosis associated with adrenal masses and insufficiency; Kawamura M et al.; A case of primary adrenal insufficiency with bilateral adrenal masses and meningitis due to disseminated cryptococcosis in a patient with mild non-insulin-dependent diabetes is presented . The diagnosis was made by fine-needle aspiration biopsy cytology . Although the meningitis responded to antifungal therapy, the bilateral adrenal gland enlargement did not change . Reflecting this, cryptococcal antigen titers became negative in CSF, but fell to 1:8 in serum . Although antifungal therapy continued, cryptococcal antigen titer increased both in CSF and serum for 50 days . Because the adrenal glands were the apparent focus for the persistent fungemia, bilateral adrenalectomy was performed . Antifungal therapy for an additional 15 months was needed to achieve negative serum cryptococcal antigen titers . Although adrenal insufficiency due to disseminated cryptococcosis is rare in healthy hosts, it should be included in differential diagnosis of unilateral and bilateral adrenal masses.

Mycoses, 1998 Mar-Apr, 41(3-4), 117 - 24
Molecular subtyping of clinical and environmental strains of Cryptococcus neoformans variety neoformans serotype A isolated from southern Italy; Pernice I et al.; Analysis of ribosomal DNA (rDNA) restriction fragment-length polymorphism (RFLP) and random amplification of polymorphic DNA (RAPD) was used to investigate the genetic variability and biogeographic distribution of clinical and environmental strains of Cryptococcus neoformans isolated from a limited area of southern Italy, where the selection of a predominant cryptococcal genotype could be expected . All isolates belonged to the species Cr . neoformans variety neoformans serotype A . RFLP analysis of a specific rDNA fragment allowed the distinction of strains of Cr . neoformans from closely related fungal reference species, but neither intraspecies nor intravarieties polymorphism was detected . On the contrary, RAPD fingerprints produced by priming with four different primers {(GTG)5, (GACA)4, M13 core sequence and the 8-mer oligonucleotide (GCGGACGG)} were able to characterize the isolates up to the individual level, indicating the presence of marked heterogeneity among Cr . neoformans serotype A strains in southern Italy.

Curr Opin Chem Biol, 1997 Aug, 1(2), 176 - 82
The search for new triazole antifungal agents; Koltin Y et al.; The first generation antifungal agent triazoles, fluconazole and itraconazole, have revolutionised the treatment of serious fungal infections such as mucosal and invasive candidiasis and cryptococcal meningitis . However, the treatment of some fungal infections, particularly aspergillosis, is still far from satisfactory and thus there is an important requirement for new broad-spectrum antifungal agents . The new second generation triazoles voriconazole and SCH-56592 show considerable promise in achieving this goal in the near future.

J Clin Microbiol, 1998 Aug, 36(8), 2200 - 4
Genetic relationship between Cryptococcus neoformans var . neoformans strains of serotypes A and D; Franzot SP et al.; Cryptococcus neoformans serotypes A and D are responsible for the overwhelming majority of infections in patients with AIDS . The genetic relationship between the serotypes is poorly understood, but there are significant differences in the epidemiology and clinical presentation of serotype A and D infections . We evaluated the genetic relationship between reference C . neoformans strains belonging to serotypes A and D by analyzing their URA5 sequences and restriction fragment length polymorphisms (RFLPs) with the C . neoformans repetitive element 1 (CNRE-1) probe . The results were compared to those previously obtained for isolates from Brazil and New York City by the same typing methods, and dendrograms were generated . Serotype A and D strains produced distinct RFLP patterns consistent with their separation into two major clusters in the dendrogram generated on the basis of RFLP data . Similarly, serotype A and D strains clustered independently of the basis of the nucleotide sequences of their URA5 genes . Pairwise comparisons revealed average numbers of nucleotide differences within serotypes A and D of 3.0 +/- 1.7 and 7.2 +/- 3.4, respectively (P < 0.0001), and between serotypes A and D of 41.9 +/- 2.7 . In summary, our results indicate phylogenetic differences between the two serotypes of C . neoformans var . neoformans and suggest that these serotypes could probably be considered different varieties of C . neoformans.

Antimicrob Agents Chemother, 1998 Jul, 42(7), 1722 - 5
Efficacy of NS-718, a novel lipid nanosphere-encapsulated amphotericin B, against Cryptococcus neoformans; Hossain MA et al.; In vitro and in vivo efficacies of NS-718, a lipid nanosphere-encapsulated amphotericin B (AMPH-B), have been studied . Of the tested AMPH-B formulations, NS-718 had the lowest MIC for Cryptococcus neoformans . In a murine model, low-dose therapy (0.8 mg/kg of body weight) with NS-718 showed higher efficacy than that with AmBisome . High-dose therapy (2.0 mg/kg) with NS-718 was much more effective than those with Fungizone and AmBisome . In mice treated with a high dose of NS-718, only a few yeast cells had grown in lung by 7 days after inoculation . A pharmacokinetic study showed higher concentrations of AMPH-B in lung following administration of NS-718 than after administration of AmBisome . Our results indicated that NS-718, a new AMPH-B formulation, is a promising antifungal agent for treatment of pulmonary cryptococcosis and could be the most effective antifungal agent against C . neoformans infections.

Biochim Biophys Acta, 1998 Jun 5, 1381(1), 61 - 7
Effects of Cryptococcus humicola killer toxin upon Cryptococcus terreus envelope: combined fluorometric and microscopic studies; Puchkov EO et al.; Killer toxin (microcin) produced by Cryptococcus humicola 9-6 induced interaction of the fluorogenic dyes, ethidium bromide, propidium iodide, and hemimagnesium 8-anilino-1-naphtalenesulfonate, with the sensitive strain of Cryptococcus terreus VKM Y-2253 . The toxin also made the cells susceptible to cetyltrimethylammonium bromide and leaky for K+ . When excited at 360 nm, cell-bound ethidium (propidium) fluorescence was enhanced by 8-anilino-1-naphtalensulfonate, and cell-bound 8-anilino-1-naphtalensulfonate fluorescence was quenched by ethidium (propidium), indicating energy transfer from 8-anilino-1-naphtalensulfonate to ethidium (propidium) . These results suggest that at least a portion of the probe molecules had the same binding site, possibly the cytoplasmic membrane . The parameters of kinetics of microcin action were evaluated fluorometrically . They were found to be identical for all probes and depended on microcin concentration . The fluorescence increment of ethidium and 8-anilino-1-naphtalensulfonate upon binding to microcin-treated cells correlated with the fraction of stainable cells and viability.

J Clin Pathol, 1998 Mar, 51(3), 246 - 8
A pseudo-cryptococcal artefact derived from leucocytes in wet India ink mounts of centrifuged cerebrospinal fluid; Thiruchelvan N et al.; Wet India ink mounts of cerebrospinal fluid (CSF) are useful in the laboratory diagnosis of cryptococcal meningitis . Pseudo-cryptococcal artefacts in such mounts have been attributed to leucocytes in CSF but their mode of formation has not been explained . This report describes the reproduction of such an artefact in cryptococcus free CSF-leucocyte mixtures that had been subjected to high speed centrifugation . The viscosity of DNA that could provide a morphological pseudo-capsule, and the yellow-green fluorescence of the pseudo-capsular material on staining with acridine-orange, suggest that lymphocytic nuclear DNA, which possibly leaked out after damage to the lymphocyte membrane by centrifugation, was responsible for this artefact.

Nihon Kokyuki Gakkai Zasshi, 1998 Mar, 36(3), 299 - 305
{Cytokines produced by cells in bronchoalveolar lavage fluid from a patient with primary pulmonary cryptococcosis}; Abe K et al.; Cytokines in the culture supernatant of concanavalin A-stimulated macrophages/lymphocytes isolated from bronchoalveolar lavage (BAL) fluid in a 29-year-old patient with primary pulmonary cryptococcosis were evaluated to study the immune reaction against Cryptococcus neoformans in the lung . Before fungicidal therapy, levels of interferon-gamma and interleukin-10 were markedly elevated, and declined after therapy . There were no changes in interleukin-2 or interleukin-4 throughout the clinical course . This result suggests that IFN-gamma and IL-10 may be involved in the immune reaction against pulmonary cryptococcosis.

Nihon Kokyuki Gakkai Zasshi, 1998 Mar, 36(3), 256 - 61
{A case of Wegener's granulomatosis which showed early spontaneous remission}; Yanagawa T et al.; An 80-year-old woman presented at our hospital on October 1995 with fever, hemoptysis and a cavitary shadow on chest X-ray . Blood examination revealed an accelerated erythrocyte sedimentation ratio and elevated CRP . Pulmonary cryptococcosis was suspected, but serological tests and bronchoscopic examination for cryptococcus were both negative . There was also no evidence of the tuberculosis or malignancy . She was treated with the antibiotic cefpirome sulfate intravenously for thirteen days . Her chest X-ray and abnormal blood test findings became almost completely normal following the i.v . antibiotic treatment . In February 1996 (2 months after her first admission), she had severe right cheek pain, and Coldwell Luc's operation was performed after right maxillary sinusitis was diagnosed . A high fever (39 degrees C) continued after surgery, and multiple cavitary shadows were seen on chest X-ray . Blood examination revealed an accelerated ESR, elevated CRP and slightly elevated c-ANCA . She was treated with i.v . infusion of antibiotics and antifungal drug's, but did not improve . Wegener's granulomatosis was diagnosed after transcutaneous lung biopsy and histopathological examination of the maxillary sinus . Dramatic improvement was seen following treatment with oral cyclophosphamide and prednisolone . Whether her first remission was due to antibiotic treatment or spontaneous is an interesting question.

Pediatr Dermatol, 1998 May-Jun, 15(3), 222 - 4
Solitary giant molluscum contagiosum of the sole; Ha SJ et al.; Molluscum contagiosum of the sole is extremely rare and only three cases have been reported in the literature . We report a solitary giant molluscum contagiosum on the left sole of a 5-year-old boy, which should be clinically differentiated from plantar wart, eccrine poroma, epidermal cyst, foreign body granuloma, cryptococcal infection, and pyogenic granuloma.

Medicine (Baltimore), 1998 May, 77(3), 153 - 67
Cryptococcal pneumonia complicating pregnancy; Ely EW et al.; In the present report we describe 4 previously healthy women who developed cryptococcal pneumonia during pregnancy, and 1 pregnant woman with cryptococcal meningitis . These cases illustrate a previously uncharacterized spectrum of cryptococcal disease . We also discuss 24 patients previously reported who had cryptococcal meningitis during pregnancy . Finally, we review the available data for each therapeutic option and present an algorithm for management based on appraisals of disease severity and risk to the unborn fetus . This report emphasizes the need for heightened awareness of cryptococcosis in the differential diagnosis of pneumonia, chest pain, and hypoxemia in the pregnant patient, but at present, there are insufficient epidemiologic data to determine whether incidences of pulmonary or disseminated cryptococcosis actually increase during pregnancy . The risk of congenital cryptococcosis to the unborn fetus is low, and the most likely mechanism whereby neonates acquire invasive fungal pulmonary infection is through aspiration . While it is unclear whether there is any real increased risk of spontaneous abortion or premature labor, the data indicate that overall fetal outcome depends on effective treatment of maternal infection . For patients with dense air-space consolidation, progressive pulmonary disease, or dissemination, antifungal therapy is necessary . Optimal treatment is determined by the acuity and severity of the clinical presentation . Amphotericin B (approximately 1 g) with or without flucytosine represents the choice for initial treatment of the more acutely ill patient with disseminated or progressive pulmonary cryptococcosis who requires hospitalization (whether during or after pregnancy) . Oral fluconazole appears to be safe and effective alternative therapy after delivery for the less severely ill patient who can be managed on an outpatient basis . While the use of fluconazole during pregnancy generally appears safe in terms of fetal outcome (49, 58), the class C status and single report of fetal malformation (62) preclude confident recommendation for its use during pregnancy . The risks and benefits of this effective and generally less toxic drug should be discussed with the parents and weighed against the use of amphotericin B . For pregnant women with limited pulmonary cryptococcosis (segmental or nodular infiltrates) and no evidence of dissemination, we recommend close follow-up without antifungal therapy similar to the recommendation for normal hosts with minimal disease . However, it is important to note that there is no extensive experience upon which to base this recommendation for pregnant individuals (45, 55, 103, 108) . It is prudent to use frequent physical examinations (for example, every 1-2 months), combined with chest roentgenograms and serum cryptococcal antigens to monitor progression and/or development of disease in both the mother and child for approximately 6 months postpartum . In conclusion, cryptococcosis during pregnancy presents a special challenge to the clinician . A balanced therapeutic approach holds great promise for successful maternal and fetal outcomes.

Chirurg, 1998 May, 69(5), 530 - 5
{Fungal infection}; Kaben U; Untreated dermatological mycoses are easy to diagnose . Once treatment with corticosteroids has been initiated the diagnosis of a mycosis can be difficult . In the immunosuppressed patient (AIDS patients) typical dermatological manifestations can be lacking . Systemic antimycotic therapy requires precise detection of the pathogen concerned . Mistakes still made in surgical practice are incision of a tumour in the case of tinea profunda and the extraction of nails affected by fungi . In the healthy person yeasts are transient organisms present in the mouth and intestinal tract in contrast, the mouth and intestinal tract of patients in risk make up a reservoir of candida infection that can affect the internal organs . Cryptococcosis and aspergillosis are inhaled mycoses . Factors predisposing to mycoses influence the duration and the outcome of the course of illness . The most important of these factors in surgical practice is the immunosuppression . Systemic mycoses are difficult to recognize . In many cases organ mycoses can be diagnosed by CT . Continuous investigations of diagnostic cultures and serological tests can contribute to the diagnosis . Only cryptococcosis can be ascertained early by specific antigen demonstration in the serum . For this reason continuous serological testing for cryptococci is essential in AIDS patients.

J Infect Dis, 1998 Jul, 178(1), 114 - 20
Trends in AIDS-related opportunistic infections among men who have sex with men and among injecting drug users, 1991-1996; Jones JL et al.; Incidence trends for the 13 most frequent AIDS-defining opportunistic infections (OIs) among men who have sex with men (MSM, n = 15,588) and injecting drug users (IDUs, n = 4475) were examined using data abstracted from medical records in >90 hospitals and clinics in nine US cities during 1991-1996 . Among MSM, the most frequent OIs were Mycobacterium avium complex (MAC) disease, Pneumocystis carinii pneumonia (PCP), and cytomegalovirus (CMV) retinitis; decreasing (P < or = .05) trends occurred for 11 OIs (MAC disease, PCP, CMV retinitis, Kaposi's sarcoma, esophageal candidiasis, CMV disease, extrapulmonary cryptococcosis, toxoplasmic encephalitis, tuberculosis, chronic herpes simplex, and disseminated histoplasmosis) . Among IDUs, the most frequent OIs were PCP, MAC disease, and esophageal candidiasis; decreasing trends occurred for 5 OIs (PCP, esophageal candidiasis, tuberculosis, chronic herpes simplex, and chronic cryptosporidiosis) and an increase occurred in recurrent pneumonia . The differences in trends for MSM and IDUs may be due to differences in medical care and adherence to preventive medications.

J Investig Med, 1998 Apr, 46(4), 153 - 60
Diagnostic characteristics of cerebrospinal fluid analysis for secondary menigitis in HIV-infected adults; Friedmann PD et al.; BACKGROUND: When human immunodeficiency virus (HIV)-infected patients present with neurologic problems, lumbar puncture and cerebrospinal fluid (CSF) analysis are often performed, usually to rule-out the presence of meningitis . However, the test characteristics of CSF analyses in this population are unknown . METHODS: To examine the diagnostic characteristics of CSF analyses for secondary causes of meningitis in HIV-infected adults, we performed a case-control study of 322 HIV-infected adults who received diagnostic lumbar puncture in an urban academic hospital from 1989 to 1992 . Using multivariable logistic regression analyses, we determined the independent CSF correlates of cryptococcal and other types of secondary meningitis . RESULTS: Eighty percent were men who had sex with men, 89% were white, and 85% had AIDS . Thirty-seven case-subjects had a secondary cause of meningitis though none had bacterial or tuberculous meningitis . Excluding the india ink as a candidate variable, logistic regression analysis identified 2 independent CSF correlates of secondary meningitis: the leukocyte count (odds ratio {OR} per 10 cells, 1.2; 95% confidence interval {CI}, 1.1 to 1.3), and the CSF-to-serum glucose ratio (OR per 0.10 units, 0.61; CI, 0.43 to 0.88) . The areas (+/- SD) under the receiver operating characteristic curves were 0.76 +/- 0.05 and 0.72 +/- 0.05, respectively . CONCLUSIONS: In similar clinical settings, these findings may help clinicians assess the probability of secondary meningitis in HIV-infected patients when initial CSF results first become available.

Pathologe, 1998 May, 19(3), 194 - 200
{Liver changes in AIDS . Retrospective analysis of 227 autopsies of HIV-positive patients}; Trojan A et al.; In a retrospective study of a 12-year period (1981-1992) liver histology was analyzed in 227 autopsied patients infected with the human immunodeficiency virus . Normal histology could only be documented in 29 patients (13%) . In the majority of cases (56%) uncharacteristic changes were seen such as steatosis (34%), hemosiderosis (10%) or non-specific reactive hepatitis (7%) . The finding of hepatic peliosis obtained in 4 patients was not associated with inflammatory liver changes, especially infections from Rochalimaea . Within a wide range of opportunistic infections recorded in 50 patients (22%), hepatitis caused by Cytomegalovirus (8%), Toxoplasma gondii (5%), Leishmania donovani (1%), Cryptococcus neoformans and Pneumocystis carinii (each 0.5%) was diagnosed . Among 16 cases (7%) of mycobacterial liver infections typical mycobacteria were found in two patients and atypical mycobacteria in 14 patients, respectively . In 23 patients (10%) chronic viral hepatitis, caused by HBV (7%) or HCV infections (3%), respectively, was observed . Hepatitis was typed as mild only in each 5 patients with HBV or HCV infection, whereas the remaining cases showed a transition towards cirrhosis . Two patients with HBV-associated cirrhosis developed hepatocellular carcinoma . The remaining 32 malignant liver tumors represented secondary neoplasms, including 13 cases of non-Hodgkin's lymphomas.

Mycopathologia, 1997-98, 140(2), 65 - 8
Pulmonary cryptococcosis due to a capsule-deficient strain confused with metastatic lung cancer; Kimura M et al.; A patient with hepatocellular cancer developed pulmonary cryptococcosis due to infection with a capsule-deficient Cryptococcus neoformans . Pulmonary lesions initially diagnosed as metastatic cancer by chest x-ray film and CT scan were subsequently found to be fungal granulomas by autopsy . Although morphologic studies of the fungi were insufficient to render a specific mycologic diagnosis because of the absence of encapsulated yeasts, fluorescent antibody studies confirmed the diagnosis of cryptococcosis . The use of various stains and electron microscopy for the pathological differential diagnosis of cryptococcosis caused by capsule-deficient yeasts is discussed.

Ann Ital Med Int, 1998 Jan-Mar, 13(1), 8 - 12
{AIDS-related cryptococcosis: diagnostic aspects, prognostic and therapeutic implications}; Manfredi R et al.; Diagnostic techniques of AIDS-related cryptococcosis were assessed in a series of 43 HIV-infected patients, and microbiological features were correlated to the clinical course and outcome of disease . Polysaccharide antigen detection was the most sensitive method for central nervous system infection, followed by direct microscopy and culture: in 4 patients this visceral mycosis was initially diagnosed by the detection of isolated cerebrospinal fluid antigen . Thirty-one patients out of 43 suffered from disseminated infection (with positive blood cultures and/or antigenemia) . The occurrence of clinical relapses, a lethal outcome, and time to relapses or to death, did not differ significantly between patients with isolated central nervous system and those with disseminated cryptococcosis.

Microbiology, 1998 Jun, 144 ( Pt 6), 1691 - 7
A novel quantitative mating assay for the fungal pathogen Cryptococcus neoformans provides insight into signalling pathways responding to nutrients and temperature; Dong H et al.; Cryptococcus neoformans is a fungal pathogen that causes a lethal meningitis in immunocompromised individuals . Several factors are associated with virulence of this fungus, including its mating type; however, the mechanism by which mating type affects virulence is unknown . C . neoformans is a basidiomycete that exists in two mating types called a and alpha that can fuse to form an a/alpha dikaryon . A mating assay was developed that allowed a quantitative analysis of cryptococcal mating physiology . Interestingly, the efficiency of mating appeared to be dependent on temperature, being highest at 30 degrees C and almost completely absent at 37 degrees C . Thus, while mating type itself may be associated with virulence (which must occur at 37 degrees C), the ability to mate is probably not a virulence factor . Mating efficiency was increased by altering the carbon or nitrogen sources to give so-called starvation media . The addition of various drugs also seemed to alter the frequency of mating, depending on the composition of mating medium . The data suggested that cAMP, 8-bromo-cAMP and caffeine increased mating on starvation medium but only cAMP and 8-bromo-cAMP stimulated mating on rich medium; caffeine was unable to stimulate mating on rich medium . Aluminium fluoride, an activator of heterotrimeric GTP-binding proteins (G-proteins), was also found to stimulate mating, suggesting the involvement of a G-protein that may regulate the level of cAMP.

Zentralbl Bakteriol, 1998 May, 287(4), 489 - 500
Cytomegalovirus infection in kidney transplant patients: clinical manifestations and diagnosis; Sulowicz W et al.; Cytomegalovirus (CMV) infection is an important cause of morbidity and mortality after solid organ transplantation . CMV infection after kidney transplantation was confirmed in 19 (54.3%) out of 35 patients . 16 of these (84.2%) developed CMV disease . CMV infection was diagnosed based on a fourfold or greater increase of anti-CMV IgG antibody titre, detection of CMV-IgM antibodies and/or virus isolation . Primary infection was observed in 3 patients, reactivation in 9 and an undefined type of infection in 7 . In most patients (63%), infection was diagnosed in the first 2 months, and in 3 patients, after 3, 5 and 9 years following kidney transplantation . The most frequent symptoms of CMV disease were fever (58%), pneumonitis (26.3%) and enterocolitis (15.8%) . In 53% of the patients, CMV infection co-occurred with other pathogens such as Candida albicans . Cryptococcus neoformans, bacteria or viruses (HBV, HCV, HSV) . Treatment with polyvalent globulin (Sandoglobin) or hyperimmune globulin (Cytotect), in combination with ganciclovir in 7 patients, resulted in a regression of CMV disease.

Neuroradiology, 1998 May, 40(5), 303 - 7
MRI features of choroid plexitis; Cho IC et al.; Four cases of choroid plexitis of the brain (two with cryptococcosis and two with tuberculosis) are presented . The four patients showed either unilateral enlargement (3) or bilateral enlargement (1) and dense enhancement of the choroid plexus in the lateral ventricles (4) and fourth ventricle (1) in association with clinical findings of leptomeningitis . All patients had unilateral cystic dilatation of the temporal horn of the lateral ventricle presumably secondary to entrapment of the temporal horn and extensive oedema around the ipsilateral ventricle.

Wiad Lek, 1995 Jan-Jun, 48(1-12), 234 - 41
{Central nervous system infections in patients with AIDS}; Niwicka-Michalowska A; Central Nervous System (CNS) is very common site of the opportunistic infections in patients with AIDS . Patients, who died because of AIDS have described pathology of CNS in 80% in autopsy series . Toxoplasmic encephalitis (TE) is the most common infection in the course of AIDS, and it touches 25-50% of the HIV-infected people . The treatment of TE is very difficult, but relapses are very often and primary and secondary prophylaxis of TE is necessary . Fungal infections (particularly cryptococcal meningitis) are very unpopular in immunocompetent patients; in HIV-infected people Cryptococcus neoformans is the cause of the 30% of encephalitis . Viral and bacterial encephalitis, they are not very common in AIDS patients.

Am J Respir Crit Care Med, 1998 Jun, 157(6 Pt 1), 1913 - 8
Transbronchial needle aspiration (TBNA) in patients infected with HIV; Harkin TJ et al.; Transbronchial needle aspiration (TBNA) of intrathoracic lymph nodes has been shown to be useful in the diagnosis and staging of bronchogenic carcinoma . With the exception of sarcoidosis, the usefulness of TBNA has not been widely investigated in other clinical settings . We investigated the utility of TBNA with a 19-gauge histology needle in HIV-infected patients with mediastinal and hilar adenopathy at Bellevue Hospital Center . We performed 44 procedures in 41 patients . Adequate lymph node sampling was obtained in 35 of 44 (80%), and diagnostic material was obtained in 23 of 44 (52%) procedures . TBNA was the exclusive means of diagnosis in 13 of 41 (32%) patients . Of the 44 procedures, 23 (52%) were performed in patients with mycobacterial disease, with TBNA providing the diagnosis in 20 of 23 (87%) . In these patients, positive TBNA specimens included smears of aspirated materials for acid-fast bacilli in 11, mycobacterial culture in 14, and histology in 15 . In other diseases, TBNA diagnosed sarcoidosis with noncaseating granulomata in 2 of 4 patients and non-small cell lung cancer in 1 of 2 patients . TBNA was not helpful in other diseases including Pneumocystis carinii pneumonia, infection with Cryptococcus or Nocardia, bacterial pneumonia, viral pneumonia, and Kaposi's sarcoma . No pulmonary diagnosis was established in five patients . No complications of TBNA occurred . We conclude that TBNA through the flexible bronchoscope is safe and effective in the diagnosis of intrathoracic adenopathy in HIV-infected patients, and is particularly efficacious in the diagnosis of mycobacterial disease . Furthermore, TBNA may provide the only diagnostic specimen in almost one-third of HIV-infected patients, thereby sparing these patients more invasive procedures such as mediastinoscopy.

J Immunol, 1998 Jun 15, 160(12), 6026 - 31
The antibody response to fungal melanin in mice; Nosanchuk JD et al.; Melanins are associated with virulence in several important human pathogens, but little is known about the immune response to this ubiquitous biologic compound . We hypothesized that melanin produced by the fungus Cryptococcus neoformans was immunogenic . C . neoformans melanin was purified from melanized fungal cells and was used to immunize C57BL/6, BALB/c, and T cell-deficient (nude) BALB/c mice . The Ab response was evaluated by ELISA, immunofluorescence, and agglutination . The results demonstrate that melanin can be immunogenic, and the humoral immune response is T cell independent . Furthermore, the experiments demonstrate 1) a sensitive ELISA for the measurement of Ab to melanin, 2) that mice mount an intense Ab response to fungal melanin that includes Abs of IgM and IgG isotypes, 3) that melanins from different sources have cross-reactive epitopes, and 4) melanin in the cell wall of melanized yeast cells reacts with Abs raised to L-dopa C . neoformans melanin . The biologic significance of Ab to melanin remains to be determined, but the development of Ab suggests that this amorphous insoluble polymer can stimulate the immune system . The serologic techniques described here may prove useful for the evaluation of Ab responses to melanin in a variety of diseases.

Clin Neurol Neurosurg, 1998 Mar, 100(1), 51 - 2
Reversible blindness in AIDS-related cryptococcal meningitis; Claus JJ et al.; A 30-year-old AIDS-patient with cryptococcal meningitis developed subacute bilateral visual loss associated with high cerebrospinal fluid (CSF) pressure . With immediate CSF drainage the blindness was reversible . The importance of prompt CSF drainage in AIDS-related cryptococcal meningitis with visual failure is stressed.

Clin Infect Dis, 1998 Jun, 26(6), 1362 - 6
Combination therapy with fluconazole and flucytosine for cryptococcal meningitis in Ugandan patients with AIDS; Mayanja-Kizza H et al.; We performed a randomized trial in which combination therapy with fluconazole and short-term flucytosine was compared with fluconazole monotherapy in 58 patients with AIDS-associated cryptococcal meningitis (CM) . Thirty of these patients were randomized to receive combination therapy with fluconazole, 200 mg once a day for 2 months, and flucytosine, 150 mg/(kg.d) for the first 2 weeks, and 28 were randomized to receive monotherapy with fluconazole at the same dose for 2 months . Patients in both groups who survived for 2 months received fluconazole as maintenance therapy at a dose of 200 mg three times per week for 4 months . The combination therapy prevented death within 2 weeks and significantly increased the survival rate among these patients (32%) at 6 months over that among patients receiving monotherapy (12%) (P = .022) . The combination therapy also resulted in a significant decrease in the severity of headache after 1 month of treatment, compared with monotherapy (P = .005) . No serious adverse reactions were observed in patients receiving either regimen . These data indicate that treatment with fluconazole and short-term flucytosine is a cost-effective and safe regimen that improves the quality of life for patients with AIDS-associated CM in developing countries where human immunodeficiency virus is endemic.

Allergy, 1998 May, 53(5), 506 - 12
IgE, IgA, and IgG responses to common yeasts in atopic patients; Savolainen J et al.; This study was undertaken to analyze the differences in exposure and sensitization to five common environmental yeasts . The responses of IgG, IgA, and IgE to Candida albicans, C . utilis, Cryptococcus albidus, Rhodotorula rubra, and Saccharomyces cerevisiae and purified S . cerevisiae enolase were analyzed by immunoblotting (IgE-IB), and the cross-reactivity of their IgE-binding components by IgE-IB inhibition . Twenty atopic subjects, with asthma, allergic rhinitis, or atopic dermatitis were included . In skin prick tests (SPT), 12 of the patients showed simultaneous reactivity to at least two of the five yeasts, four reacted to one of the yeasts, and four had no responses . Antigens run in SDS-PAGE and transferred to nitrocellulose were probed with enzyme-labeled IgA-, IgG-, and IgE-specific antibodies . The IgE immunoblotting revealed most IgE-binding bands in C . albicans (11 bands) followed by C . utilis (eight bands), S . cerevisiae (five bands), R . rubra (five bands), and Cr . albidus (four bands) . Six of the IgE-binding bands of C . albicans and C . utilis shared molecular weight, and only two bands shared molecular weight with other yeasts . These were the 46-kDa band, shared by all five yeasts, and a 13-kDa band shared by four yeasts . Prominent IgE binding was seen to a 46-kDa band of C . albicans (seven patients), C . utilis (five patients), and S . cerevisiae (one patient) and to corresponding weak bands of Cr . albidus and R . rubra (one patient) . The possible cross-reactivity of the 46-kDa band was analyzed by IgE-IB inhibition and densitometry, revealing clear C . albicans inhibition of C . utilis (80%) and enolase (98%) (autoinhibition 100%) . The strongest IgG responses were seen against S . cerevisiae and C . albicans . The responses were mainly against mannans of C . albicans and S . cerevisiae, suggesting that most of the exposure is to these yeasts . Yeasts with different types of exposure, from saprophytic growth on human mucous membranes to exposure by air and food, were shown to cross-react at the allergenic level . Atopic patients primarily sensitized by C . albicans and S . cerevisiae may develop allergic symptoms by exposure to other environmental yeasts due to cross-reacting IgE antibodies.

Neuropathol Appl Neurobiol, 1998 Apr, 24(2), 118 - 24
HIV-associated brain pathology: a comparative international study; Davies J et al.; Little is known about the frequency and variation of HIV-associated brain pathology in different geographical centres . To assess whether there is an association between the frequency of disease and demographic factors we examined the neuropathological findings in four European and two American cities . The cities included London, Edinburgh, Paris, Budapest, Baltimore and Newark . Information was collected on a total of 1144 cases . HIV encephalitis was the most common observation in all the centres . although its frequency varied between them (P < 0.01) . Furthermore, there were significant differences (P < 0.001) between the various categories of exposure and the frequency of HIV encephalitis in Edinburgh and other centres . The occurrence of toxoplasmosis, progressive multifocal leukoencephalolpathy (PML) and cryptococcal infection also differed between the various centres (P < 0.01) . None of the findings was attributable to age, sex, or ethnic origin, but the introduction of anti-retroviral treatment, such as Zidovudine, may have been important . Overall, this study highlights geographical variability and the potential importance for group of exposure and anti-retroviral medication as factors affecting the development of various HIV-associated brain lesions.

J Biol Chem, 1998 May 15, 273(20), 12482 - 91
Genetic and biochemical studies establish that the fungicidal effect of a fully depeptidized inhibitor of Cryptococcus neoformans myristoyl-CoA:protein N-myristoyltransferase (Nmt) is Nmt-dependent; Lodge JK et al.; Cryptococcus neoformans is a fungal pathogen that causes chronic meningitis in 10% of patients with AIDS . Genetic and biochemical studies were conducted to determine whether myristoyl-CoA:protein N-myristoyltransferase (Nmt) is a target for development of a new class of fungicidal drugs . A single copy of a conditional lethal C . neoformans NMT allele was introduced into the fungal genome by homologous recombination . The allele (nmt487D) produces temperature-sensitive myristic acid auxotrophy . This phenotype is due, in part, to under-myristoylation of a cellular ADP ribosylation factor (Arf) and can be rescued by forced expression of human Nmt . Two isogenic strains with identical growth kinetics at 35 degreesC were used to test the biological effects of an Nmt inhibitor . CPA8 contained a single copy of wild type C . neoformans NMT . HMC1 contained nmt487D plus 10 copies of human NMT . Since a single copy of nmt487D will not support growth at 35 degreesC, survival of HMC1 depends upon its human Nmt . ALYASKLS-NH2, an inhibitor derived from an Arf, was fully depeptidized: p-{(2-methyl-1-imidazol-1-yl)butyl}phenyl-acetyl was used to represent the GLYA tetrapeptide, whereas SKLS was replaced with a chiral tyrosinol scaffold . Kinetic studies revealed Ki (app) values of 1.8 +/- 1 and 9 +/- 2.4 microM for purified fungal and human Nmts, respectively . The minimal inhibitory concentration of the compound was 2-fold lower for CPA8 compared with HMC1 . A single dose of 100 microM produced a 5-fold greater inhibition of protein synthesis in CPA8 versus HMC1 . The strain specificity of these responses indicates that the fungicidal effect was Nmt-dependent . These two strains may be useful for screening chemical libraries for Nmt-based fungicidal compounds with relatively little activity against the human enzyme.

J Am Acad Dermatol, 1998 Jun, 38(6 Pt 2), S95 - 102
Once-weekly fluconazole (450 mg) for 4, 6, or 9 months of treatment for distal subungual onychomycosis of the toenail; Ling MR et al.; BACKGROUND: Fluconazole is a bis-triazole antifungal agent approved for the treatment of oropharyngeal, esophageal, and vaginal candidiasis, serious systemic candidal infections, and cryptococcal meningitis . OBJECTIVE: The purpose of this study was to evaluate three different durations of once-weekly fluconazole for the treatment of onychomycosis of the toenail caused by dermatophytes . METHODS: In a multicenter, randomized, double-blind, parallel, placebo-controlled trial, 384 patients with distal subungual onychomycosis of the toenail received fluconazole, 450 mg once weekly, or placebo for 4, 6, or 9 months . For inclusion, patients were required to have mycologically confirmed distal subungual onychomycosis of the toenail with a large toenail at least 25% clinically affected but having at least 2 mm of healthy nail between the nail fold and the proximal onychomycotic border . Efficacy was assessed by clinical and mycologic (microscopic and microbiologic) measures at screening, at every treatment visit starting at month 3, and at months 2, 4, and 6 after therapy . Observed or volunteered adverse events were recorded and classified at all visits . RESULTS: At the end of treatment, very significantly superior clinical and mycologic results were achieved in all fluconazole groups compared with placebo (p=0.0001) . This superiority was largely maintained over 6 months of follow-up . The clinical and mycologic responses of the 9-month treatment duration were significantly superior to the 4- and 6-month durations . Similar percentages of patients in the fluconazole and placebo groups reported adverse experiences for all three durations of the study . CONCLUSION: Results of this study support the efficacy and safety of fluconazole in the treatment of distal subungual onychomycosis of the toenail.

Neurol Neurochir Pol, 1998 Jan-Feb, 32(1), 155 - 60
{Diagnostic difficulties in a case of meningo-cerebral primary cryptococcosis}; Taraszewska A et al.; A case of meningo-cerebral cryptococcosis is reported in a 34-year old women, in which no evidence of associated disease predisposing to disseminated infection could be demonstrated by clinical and autopsy examination . Diagnostic difficulties and presumable factors contributing to development of disease in the cases of so-called primary cryptococcosis are discussed.

J Biol Chem, 1998 Jun 12, 273(24), 14942 - 9
Rustmicin, a potent antifungal agent, inhibits sphingolipid synthesis at inositol phosphoceramide synthase; Mandala SM et al.; Rustmicin is a 14-membered macrolide previously identified as an inhibitor of plant pathogenic fungi by a mechanism that was not defined . We discovered that rustmicin inhibits inositol phosphoceramide synthase, resulting in the accumulation of ceramide and the loss of all of the complex sphingolipids . Rustmicin has potent fungicidal activity against clinically important human pathogens that is correlated with its sphingolipid inhibition . It is especially potent against Cryptococcus neoformans, where it inhibits growth and sphingolipid synthesis at concentrations <1 ng/ml and inhibits the enzyme with an IC50 of 70 pM . This inhibition of the membrane-bound enzyme is reversible; moreover, rustmicin is nearly equipotent against the solubilized enzyme . Rustmicin was efficacious in a mouse model for cryptococcosis, but it was less active than predicted from its in vitro potency against this pathogen . Stability and drug efflux were identified as two factors limiting rustmicin's activity . In the presence of serum, rustmicin rapidly epimerizes at the C-2 position and is converted to a gamma-lactone, a product that is devoid of activity . Rustmicin was also found to be a remarkably good substrate for the Saccharomyces cerevisiae multidrug efflux pump encoded by PDR5.

Antimicrob Agents Chemother, 1998 Jun, 42(6), 1437 - 46
Characterization of a murine monoclonal antibody to Cryptococcus neoformans polysaccharide that is a candidate for human therapeutic studies; Casadevall A et al.; The murine monoclonal antibody (MAb) 18B7 {immunoglobulin G1(kappa)} is in preclinical development for treatment of Cryptococcus neoformans infections . In anticipation of its use in humans, we defined the serological and biological properties of MAb 18B7 in detail . Structural comparison to the related protective MAb 2H1 revealed conservation of the antigen binding site despite several amino acid differences . MAb 18B7 was shown by immunofluorescence and agglutination studies to bind to all four serotypes of C . neoformans, opsonize C . neoformans serotypes A and D, enhance human and mouse effector cell antifungal activity, and activate the complement pathway leading to deposition of complement component 3 (C3) on the cryptococcal capsule . Administration of MAb 18B7 to mice led to rapid clearance of serum cryptococcal antigen and deposition in the liver and spleen . Immunohistochemical studies revealed that MAb 18B7 bound to capsular glucuronoxylomannan in infected mouse tissues . No reactivity of MAb 18B7 with normal human, rat, or mouse tissues was detected . The results show that both the variable and constant regions of MAb 18B7 are biologically functional and support the use of this MAb in human therapeutic trials.

Antimicrob Agents Chemother, 1998 Jun, 42(6), 1412 - 6
Comparison of in vitro antifungal activities of free and liposome-encapsulated nystatin with those of four amphotericin B formulations; Johnson EM et al.; The in vitro activity of a multilamellar liposomal formulation of nystatin (Nyotran) was compared with those of free nystatin and four pharmaceutical preparations of amphotericin B . MICs for 200 isolates of two Aspergillus spp., seven Candida spp., and Cryptococcus neoformans were determined by a broth microdilution adaptation of the method recommended by the National Committee for Clinical Laboratory Standards . Minimum lethal concentrations (MLCs) of the six antifungal preparations were also determined . Both nystatin formulations possessed fungistatic and fungicidal activities against the 10 species tested . Liposomal nystatin appeared to be as active as free nystatin, with MICs and MLCs that were similar to, or lower than, those of the latter . Neither formulation of nystatin was as active as amphotericin B deoxycholate (Fungizone) or amphotericin B lipid complex (Abelcet), but both were more effective than liposomal amphotericin B (AmBisome) . Our results suggest that further evaluation of liposomal nystatin is justified.

Kansenshogaku Zasshi, 1998 Apr, 72(4), 352 - 7
{A clinical study of pulmonary cryptococcosis . The Study Group of Respiratory Mycosis in Kyoto}; Kurasawa T et al.; During the 7 years from 1990, thirty-two patients (20 in male and 12 in female, mean age; 53 years old) were diagnosed as having pulmonary cryptococcosis . To clarify the essential points for early diagnosis of pulmonary cryptococcosis, we reviewed the clinical records and chest images . Three patients had a past history of pulmonary tuberculosis and eleven patients had underlying disorders such as malignancy, chronic pulmonary diseases and so on, but no HIV infection, which would affect this disease . Eighteen patients did not have any past history nor complications . The symptoms such as cough, sputum, chest pain and fever were generally of low-grade, 14 patients had no symptom at diagnosis . Except of some patients with severe infections and severe underlying disorders, laboratory findings such as inflamatory and nutritious markers were almost within near the normal range . On plain chest X-ray films the distribution of lesions was almost in proprtion to the volume of the lobes . The multifocal nudular and/or infitrative shadows wer observed in about 2/3 cases and single lesion in about 1/3 . The width of lesions were minimal except of one case with interstitial pneumonia and two cases with multifocal segmental pneumonia . The cavity lesions were observed in 7 cases and hilar lymphadenopathy in 3 cases . On CT images, the lesions were almost located in the outer zone, the lesions which were adjacent to the pleura were observed in 15 cases . Cavitary lesions were almost smooth in edge and ubiquitous, the walls were also thick . The peripheral air-bronchogram in the nodular/infitrative shadows were observed in three cases . Pulmonary cryptococcosis is air-borne and almost a chronic infection except in AIDS patients, so careful planning for examination is essential with considerations of the characteristics of clinical and imaging features of this infection.

J Clin Microbiol, 1998 Jun, 36(6), 1617 - 20
Microscopic examination and broth culture of cerebrospinal fluid in diagnosis of meningitis; Dunbar SA et al.; We reviewed the results of microscopic Gram stain examination and routine culture for 2,635 cerebrospinal fluid (CSF) samples processed in an adult hospital microbiology laboratory during 55 months . There were 56 instances of bacterial or fungal meningitis (16 associated with central nervous system {CNS} shunt infection), four infections adjacent to the subarachnoid space, four cases of sepsis without meningitis, and an additional 220 CSF specimens with positive cultures in which the organism isolated was judged to be a contaminant . Because 121 of these contaminants were isolated in broth only, elimination of the broth culture would decrease unnecessary work . However, 25% of the meningitis associated with CNS shunts would have been missed by this practice . The most common cause of meningitis was Cryptococcus neoformans, followed by Streptococcus pneumoniae and Neisseria meningitidis . In 48 of 56 (88%) of cases, examination of the Gram-stained specimen revealed the causative organism . If patients who had received effective antimicrobial therapy prior to lumbar puncture are excluded, the CSF Gram stain is 92% sensitive . Microscopic examination incorrectly suggested the presence of organisms in only 3 of 2,635 (0.1%) CSF examinations . Thus, microscopic examination of Gram-stained, concentrated CSF is highly sensitive and specific in early diagnosis of bacterial or fungal meningitis.

J Clin Microbiol, 1998 Jun, 36(6), 1625 - 9
Rapid extraction of genomic DNA from medically important yeasts and filamentous fungi by high-speed cell disruption; Muller FM et al.; Current methods of DNA extraction from different fungal pathogens are often time-consuming and require the use of toxic chemicals . DNA isolation from some fungal organisms is difficult due to cell walls or capsules that are not readily susceptible to lysis . We therefore investigated a new and rapid DNA isolation method using high-speed cell disruption (HSCD) incorporating chaotropic reagents and lysing matrices in comparison to standard phenol-chloroform (PC) extraction protocols for isolation of DNA from three medically important yeasts (Candida albicans, Cryptococcus neoformans, and Trichosporon beigelii) and two filamentous fungi (Aspergillus fumigatus and Fusarium solani) . Additional extractions by HSCD were performed on Saccharomyces cerevisiae, Pseudallescheria boydii, and Rhizopus arrhizus . Two different inocula (10(8) and 10(7) CFU) were compared for optimization of obtained yields . The entire extraction procedure was performed on as many as 12 samples within 1 h compared to 6 h for PC extraction . In comparison to the PC procedure, HSCD DNA extraction demonstrated significantly greater yields for 10(8) CFU of C . albicans, T . beigelii, A . fumigatus, and F . solani (P < or = 0.005), 10(7) CFU of C . neoformans (P < or = 0.05), and 10(7) CFU of A . fumigatus (P < or = 0.01) . Yields were within the same range for 10(8) CFU of C . neoformans and l0(7) CFU of C . albicans for both HSCD extraction and PC extraction . For 10(7) CFU of T . beigelii, PC extraction resulted in a greater yield than did HSCD (P < or = 0.05) . Yields obtained from 10(8) and 10(7) CFU were significantly greater for filamentous fungi than for yeasts by the HSCD extraction procedure (P < 0.0001) . By the PC extraction procedure, differences were not significant . For all eight organisms, the rapid extraction procedure resulted in good yield, integrity, and quality of DNA as demonstrated by restriction fragment length polymorphism, PCR, and random amplified polymorphic DNA . We conclude that mechanical disruption of fungal cells by HSCD is a safe, rapid, and efficient procedure for extracting genomic DNA from medically important yeasts and especially from filamentous fungi.

J Neurol Sci, 1998 May 7, 157(2), 201 - 5
Late onset immunodeficiency in a patient with recurrent thymic carcinoma and myasthenia gravis; Schmidt S et al.; The most common autoimmune disease associated with thymoma is myasthenia gravis . In addition, cellular and humoral immune defects have been frequently reported in association with thymic neoplasms . Here we report the case of a patient with myasthenia gravis receiving long-term immunosuppression with azathioprine and recurrent well-differentiated thymic carcinoma who developed CD4+ T-cell depletion and CNS cryptococcosis after multiple courses of chemotherapy and mediastinal irradiation . We hypothesize that in thymectomized patients bone marrow suppression and abrogation of the peripheral T-cell pool can result in a delayed T-cell regeneration due to the lack of functional thymic epithelium.

J Immunol Methods, 1998 Feb 1, 211(1-2), 51 - 63
Flow cytometric quantitation of yeast a novel technique for use in animal model work and in vitro immunologic assays; Chang WL et al.; Animal models of fungal and other infectious diseases often require that the number of organisms in tissue be quantified, traditionally by grinding organs, plating them on agar and counting colony forming units (CFU) . This method is labor intensive, slow as some fungi require two weeks of culture and limited in reliability by poor plating efficiency . To circumvent these problems, we developed a flow cytometric method to quantify yeast . In vitro cultured Blastomyces dermatitidis, Cryptococcus neoformans, Candida albicans and Histoplasma capsulatum yeast were labelled with specific monoclonal or polyclonal antibodies to stain surface determinants or with Calcofluor to stain cell-wall chitin . A defined number of fluorescently labelled beads were added prior to acquisition by flow cytometry as a reference standard for quantitation . Beads were readily distinguished from yeast by forward scatter, side scatter and intensity of fluorescence . Cultured yeast were enumerated by both standard CFU determination and flow cytometry in a range of 10(2) to 10(7) cells . Only flow cytometry enabled discrimination of live and dead yeast by using appropriate fluorescent dyes . The flow cytometric method was applied to murine models of histoplasmosis and blastomycosis to quantify the burden of fungi in the lungs of infected mice . Labelling yeast with Calcofluor alone resulted in unacceptably high levels of nonspecific binding to mouse cell debris . In contrast, labelling H . capsulatum with a rabbit polyclonal antiserum and B . dermatitidis with a monoclonal antibody to the surface protein WI-1 permitted accurate quantitation . We conclude that this flow cytometry technique is rapid, efficient and reliable for quantifying the burden of infection in animal models of fungal disease . The technique also should lend itself to performing cytotoxicity assays that require discrimination of live and dead fungi, or phagocytosis assays that require discrimination of intracellular and extracellular organisms.

Clin Immunol Immunopathol, 1998 May, 87(2), 163 - 7
Morphine enhances complement receptor-mediated phagocytosis of Cryptococcus neoformans by human microglia; Lipovsky MM et al.; Recent studies have shown that opiates modulate the function of microglia, the resident macrophages of the brain . In this study, the effect of morphine on phagocytosis by human fetal microglial cells of the opportunistic fungus Cryptococcus neoformans was studied . Contrary to earlier findings with swine microglia, opsonization was required for the phagocytosis of C . neoformans by human microglia . Moreover, morphine (10(-8)M) was shown to augment uptake of opsonized C . neoformans by over 50% . This contrasts with the earlier finding of morphine-induced inhibition of phagocytosis of nonopsonized cryptococci by swine microglia . The effect of morphine on cryptococcal phagocytosis by human microglia was reversed by treatment of microglial cells with mu opiate receptor antagonists as well as by addition of anti-complement receptor antibodies to the cell cultures, indicating that both the mu opiate receptor and the complement receptor are involved in morphine-enhanced phagocytosis . These findings support the concept of opiates as neuroimmunomodulatory agents and demonstrate that the effects of opiates on microglial cells may be influenced by the animal species from which the cells are derived.

Chemotherapy, 1998 May-Jun, 44(3), 206 - 14
Role of fluconazole in the management of AIDS-related cryptococcosis, according to daily dosing; Manfredi R et al.; No unanimous consent has been reached about treatment guidelines of cryptococcosis in the setting of AIDS, as well as about optimal fluconazole dosing in both initial and suppressive therapy . In order to evaluate the relationship between fluconazole dosing and clinical and microbiological outcome of AIDS-related cryptococcosis, a retrospective study was carried out on 30 consecutive patients . Among the 12 subjects treated with fluconazole doses < 400 mg/day, an unfavorable course was significantly more frequent (early mortality, poor clinical and microbiological response, appearance of early relapses) compared with the 18 patients who received daily doses > or = 400 mg, while no differences were observed between the two treatment groups according to known risk factors for a poor prognosis . When assessing maintenance treatment (22 evaluable cases), the 15 patients receiving oral fluconazole at doses < 200 mg/day showed earlier disease relapse and mortality as opposed to the 7 individuals treated with high-dose fluconazole (> or = 200 mg/day), in the absence of significantly different risk factors for disease recurrence . Our experience pointed out a significant difference in clinical activity of fluconazole in AIDS-related cryptococcosis according to its daily dosing, and suggested 400 and 200 mg as the threshold daily dose for an effective initial and suppressive therapy, respectively, since the probability of treatment failure seemed greater with low-dose drug administration, after controlling data for prognostic markers of disease severity . Controlled studies are warranted, comparing high-dose fluconazole with standard regimens containing amphotericin B in the treatment of AIDS-associated cryptococcosis, and identifying the best fluconazole dosing for both acute-phase and maintenance treatment.

Leuk Lymphoma, 1998 Feb, 28(5-6), 617 - 20
Cryptococcal meningitis and intracranial tuberculoma in a patient with Waldenstrom's macroglobulinemia treated with fludarabine; Costa P et al.; We report a patient with Waldenstrom's Macroglobulinemia who presented with cryptococcal meningitis followed by an intracranial tuberculoma during the 18 months period after termination of cytotoxic therapy with Fludarabine . Opportunistic infections due to intracellular organisms are extremely rare in the course of this malignancy and we review the predisposing factors of these infectious entities.

Rev Soc Bras Med Trop, 1998 May-Jun, 31(3), 257 - 61
{Adrenal gland morphological alterations in the acquired immunodeficiency syndrome}; Duch FM et al.; In the acquired immunodeficiency syndrome (AIDS), the adrenal glands are subject to opportunistic infections, neoplasm or direct cytopathic effect by HIV . It is know that the incidence and type of adrenal involvement vary according to the patient's place of origin . In this paper we evaluate adrenal involvement in fourteen patients that died from AIDS in the University Hospital of Uberaba, Brazil . The group studied was comprised of thirteen males and thirteen whites . The age was 29.9 +/- 7.8 years, and the body mass index was 19.0 +/- 4.1 kg/m2 . Adrenal specimens obtained from autopsies were analyzed by light microscopy . Inflammation was found in 100% of the cases and the etiologic agent(s) was (were) identified in eight (58.1%) patients . Cytomegalovirus was identified in seven cases, Cryptococcus sp and Herpes simplex in two and Histoplasma sp in one case, these pathologic findings were similar to literature . We also found parenchymal calcification and adrenal central vein phlebitis in one case each . Injury was found in some cases without identified infections agent . This fact could be due to the direct cytopathic effect by HIV, or due to toxicity of drug therapy used during treatment of AIDS and opportunistic infections.

Mycoses, 1998 Jan-Feb, 41(1-2), 41 - 4
Susceptibility testing of Cryptococcus neoformans using the urea broth microdilution method; Nakamura Y et al.; An urea broth microdilution method to assay the susceptibility of Cryptococcus neoformans to antifungal drugs was newly developed . Using this method, urease activity of the fungus was measured instead of the viability by checking colony development . The urease activities were indicated by colour changes in optical density at 545 nm . The end point in this assay was considered as 99% inhibitory concentration . When we measured antifungal activities of the three drugs against 16 isolates of Cr . neoformans using this assay method, mean minimum-inhibitory concentrations (MICs) of fluconazole, itraconazole and terbinafine were 2.0 micrograms ml-1, 0.008 microgram ml-1 and 0.25 microgram ml-1 respectively . This assay method resulted in higher sensitivity in MICs of the three antifungal drugs than the broth microdilution method recommended by the Committee for Laboratory Standards of the Japanese Society for Medical Mycology . The results obtained using this assay method support the more effective evaluation of antifungal substances in susceptibility testing of Cr . neoformans.

Mycoses, 1998 Jan-Feb, 41(1-2), 35 - 40
Isolates of Cryptococcus neoformans serotype A and D developed on canavanine-glycine-bromthymol blue medium; Nakamura Y et al.; Two isolates of Cryptococcus neoformans serotype A and one isolate of serotype D from pigeon droppings were found to grow on canavanine-glycine-bromthymol blue (CGB) medium, when the Japanese isolates of Cr . neoformans were examined for their serotype and biochemical characteristics . The susceptibility to canavanine and the activity in assimilation of glycine were analysed on these three isolates . They were resistant to canavanine at the high concentration of 3.6 mmol l-1 and developed by assimilating the glycine even at a concentration of 7 mmol l-1 . These isolates were proved to develop well on CGB medium, which contains 0.1 mmol l-1 of canavanine and 133 mmol l-1 of glycine . Three isolates of Cr . neoformans developed on CGB medium were also confirmed to be serotype A or D by the molecular analysis.

Mycoses, 1998 Jan-Feb, 41(1-2), 31 - 4
Deep-seated fungal diseases in the South Pacific, especially in New Caledonia; Monchy D et al.; The main deep-seated fungal diseases and their encountered pathology in New Caledonia and other islands of the South Pacific are reviewed (1970-96) . Cryptococcosis is encountered in all islands of the South Pacific, Australia and Papua New Guinea, with a predominance of variety gattii, which is associated with some species of Eucalyptus . Histoplasmosis is not uncommon, and there was an epidemic in New Caledonia in 1994 among people who had visited a bat-inhabited cave . Mycetomas, in particular presenting as pale granules in tissues, are encountered in New Caledonia, Vanuatu, Papua New Guinea, Fiji and French Polynesia . Other fungal infections, such as zygomycosis, sporotrichosis (three cases) and chromomycosis (six cases) are rarely observed in New Caledonia.

Mycopathologia, 1997, 140(1), 13 - 7
Cryptococcosis associated with AIDS in the Muñiz Hospital of Buenos Aires; Bava AJ et al.; Some epidemiologic, diagnostic and immunologic aspects of 93 (75 males and 18 women) cases of cryptococcosis (CRY) associated with AIDS, hospitalized in the Muniz Hospital (MH) in Buenos Aires during 1994, were retrospectively studied . The median age (MA) of the patients under study was 28 years (17-49 y); 30 (17-49) y for males and 23 (22-34) y for women . Intravenous drug addiction (34% of patients) and homo/bisexuality in men (17%) were the most frequent risk factors for HIV infection . The MA of these groups were 27.5 (17-41) y and 34 (25-41) y, respectively . Microscopic CSF examination with India ink and the blood cultures (lysis-centrifugation) achieved the diagnosis of CRY in 67 (72%) and 16 (17%) patients, respectively . At diagnosis, the median titers for Cryptococcus neoformans capsular antigen were 1/1,000, 1/100 and 1/1 in serum, CSF and urine, respectively . Sixty five strains of C . neoformans were insolated from clinical samples; all were identified at variety neoformans employing the culture medium proposed by Salkin & Hurd and the D-proline assimilation test . At diagnosis, CD4+ lymphocytes counts were < 50/microliter in 46 patients (83.63%), and were between 51 and 200/microliter in 9 (16.4%) . The CD4+/CD8+ ratio was < 1 in all patients . Most patients were born (83%) and lived at diagnosis (96%) in Buenos Aires (Bs As) city, Bs As outskirts towns and cities located in Bs As province.

Mycopathologia, 1997, 140(1), 1 - 11
The invasive behaviour of Cryptococcus neoformans: a possibility of direct access to the central nervous system?
Gomes NG, Boni M, Primo CC.
Cryptococcocal meningoencephalitis is always considered secondary to initial lung infection . Because of the unquestionable evidence of haematogenious spread from the lungs, few publications have reported about other possible primary sites of infection or other routes to the central nervous system . This study was designed to investigate the infiltrative pattern of C . neoformans in immunocompromised mice by treatment with dexamethasone . The infection was performed by nasal instillation (30 mice) or injection into the retro-orbital space (12 mice) . From the group infected intranasally, 3 mice presented diffuse invasive fungal colonisation of the mucosa and submucosa . The histologic findings showed infiltrative growth along the periosteum, sometimes surrounded nervous endings of submucosa, invasion along the olfactory nerve and simultaneous meningeal involvement in 2 mice on the 6th and 8th day of infection . All mice infected into the retro-ocular space developed lesions containing numerous cryptococci in the local of the inoculum . Out of the main lesion we observed preferential growth along the perineural spaces with adherence to the perineurium, perivascular spaces and sometimes along aponeurosis . Simultaneous invasion of trigeminal ganglio and trigeminal branches was observed in 4 mice . These morphologic patterns suggest the hypothesis of direct infiltrative invasion of the central nervous system.

Med Clin (Barc), 1998 Mar 28, 110(11), 406 - 10
{Invasive fungal infections in liver transplant recipients: analysis of 21 cases}; Miguelez M et al.; BACKGROUND: Invasive fungal infections (IFI) are one of the most important causes of mortality in liver transplant (LT) recipients . The aim of this study was to describe the characteristics of IFI in the LT program of our institution with an special emphasis in the differences between Candida infections (CI) and that caused by other fungi (NCI) . PATIENTS AND METHODS: Retrospective analysis of the hospital charts of 21 patients who underwent a LT from February 1987 to December 1995 . The diagnosis of IFI required the histological evidence of tissue invasion or a positive culture in a tissue sample or in an usually sterile fluid . Esophageal candidiasis was not considered as IFI . Antifungal prophylaxis was performed either with nystatin or fluconazole . RESULTS: Twenty-one of 356 patients (6%) developed a total of 23 episodes of IFI . Pathogens were Candida spp . (n = 10), Aspergillus (n = 8), Zygomicetes (n = 4) and Cryptococcus (n = 1) . Fifty-seven percent of the episodes of IFI (80% of those caused by Candida and 38% of those produced by other fungi; p < 0.05) developed in the first 3 months after transplantation and only 5 episodes appeared after the sixth month . The diagnosis of IFI was done at autopsy in 6 patients (29%) . Overall, NCI (13 episodes) predominated over CI (10 episodes), being the later the cause of the 54% of the episodes in the first 178 recipients but only the 30% in the last 178 patients (p = 0.09) . No differences were found in the distribution of the risk factors amongst those patients with CI or NCI . Seventeen of the 21 patients (71%) died and 15 of these deaths (72%) were attributable to fungi; 15 patients who died either did not receive amphotericin (n = 6) or received a cumulative dose lower than 500 mg . Six patients received a cumulative dose of more than 1.5 g (mean, 3.2 g) and four of them were cured . Mortality in the nonfungal infection group was 26% (p < 0.001) . CONCLUSIONS: IFI was a rare but severe complication in our LT recipients . The relative frequency of CI was progressively decreasing during the study period, being NCI the predominant infections . Amphotericin therapy was effective only when a high cumulative dose could be administered.

J Infect Dis, 1998 Jun, 177(6), 1647 - 59
Effect of immune mechanisms on the pharmacokinetics and organ distribution of cryptococcal polysaccharide; Lendvai N et al.; Cryptococcus neoformans is an encapsulated opportunistic fungus that can cause chronic infections accompanied by high tissue levels of capsular polysaccharide (CPS) . CPS or its major component, glucuronoxylomannan (GXM), was administered to mice, and whole-body and tissue levels were measured . The role of monoclonal antibody (MAb), complement, and CD4 T cells in GXM clearance was also examined . These studies demonstrate that CPS is cleared from the blood within days but is retained in the body for weeks; that MAbs of all isotypes examined promote GXM clearance; that MAb-mediated GXM deposition in liver, but not in spleen, is Fc-dependent; that complement enhances IgM-mediated GXM sequestration in liver but not spleen; and that CD4 T cells are not necessary for serum GXM clearance . The results have important implications for the eventual use of MAbs in treatment of cryptococcosis.

J Infect Dis, 1998 Jun, 177(6), 1639 - 46
Effect of antibody to capsular polysaccharide on eosinophilic pneumonia in murine infection with Cryptococcus neoformans; Feldmesser M et al.; The effect of the murine IgG1 monoclonal antibody (MAb) 2H1, which binds to Cryptococcus neoformans glucuronoxylomannan (GXM), on pulmonary infection in immunocompetent C57Bl/6 mice was examined . C57Bl/6 mice develop eosinophilic pneumonia in response to pulmonary cryptococcal infection . Survival, organ fungus burden, serum anticapsular antibody levels, and histopathology by light and electron microscopy were studied . MAb administration prior to infection prolonged survival without reducing the number of yeast in the lung or extrapulmonary sites . Compared with uninfected mice, occasional control and MAb-treated mice produced more IgM antibody to GXM or low levels of GXM-binding IgG1, IgG2b, or IgG3 antibodies . MAb-treated mice had fewer granules per eosinophil, indicating alteration in eosinophil physiology or degranulation (or both) . Our results provide additional evidence that antibody administration can produce quantitative and qualitative changes in the inflammatory response to a pathogen.

J Pediatr Surg, 1998 May, 33(5), 771 - 3
Cryptococcal rib osteomyelitis in a pediatric patient; Raftopoulos I et al.; A case of cryptococcal rib osteomyelitis in a pediatric patient is described . Isolated cryptococcal osteomyelitis in pediatric patients is a rare entity, and only 10 cases have been reported in the literature . The radiological findings are reviewed to include chest films, nuclear bone scan, and computed tomographic imaging scan . Because of its rarity, the management of isolated cryptococcal osteomyelitis is controversial . Although antifungal antibiotics and surgery are the two therapeutic options, the treatment of cryptococcal osteomyelitis has not been standardized yet . This patient was treated successfully with limited resection of the involved rib and antifungal chemotherapy . This article describes the second case in the literature of cryptococcal rib osteomyelitis in a pediatric patient, reviews the literature of similar cases, and evaluates the current role of surgery in its treatment.

Clin Diagn Lab Immunol, 1998 May, 5(3), 410 - 1
Effect of cytokines on anticryptococcal activity of human microglial cells; Lipovsky MM et al.; The effect of selected cytokines on the antifungal activity of human microglia was studied with encapsulated and acapsular strains of Cryptococcus neoformans . None of the cytokines tested increased the fungistatic activity of microglia, suggesting that killing of cryptococci within the central nervous system is dependent on other host defense mechanisms.

Infect Immun, 1998 Jun, 66(6), 2996 - 8
Assimilation of xylose, mannose, and mannitol for synthesis of glucuronoxylomannan of Cryptococcus neoformans determined by 13C nuclear magnetic resonance spectroscopy; Cherniak R et al.; Cryptococcus neoformans NIH 409 was cultured in a defined medium containing D-{1-13C}xylose (Xyl), D-{1-13C}mannose (Man), or D-{1-13C}mannitol as the sole carbon source . The distribution of 13C in the Man, Xyl, glucuronic acid (GlcA), and O-acetyl constituents of native and de-O-acetylated glucuronoxylomannan (GXM) was determined by one-dimensional 13C nuclear magnetic resonance spectroscopy . The carbon chain of Man was incorporated intact into GXM since 13C was observed only in carbon 1 of Man, GlcA, and Xyl . The carbon chain of mannitol was incorporated intact into GXM since 13C was observed only in carbons 1 and 6 . This was expected since mannitol has an axis of symmetry . The carbon chain of Xyl was identified only in carbons 1 and 3 of Man, GlcA, and Xyl . This pattern of labeling is consistent with the assimilation of Xyl through the pentophosphate pathway.

Infect Immun, 1998 Jun, 66(6), 2632 - 9
Antigen-induced protective and nonprotective cell-mediated immune components against Cryptococcus neoformans; Murphy JW et al.; Mice immunized with two different cryptococcal antigen preparations, one a soluble culture filtrate antigen (CneF) in complete Freund's adjuvant (CFA) and the other heat-killed Cryptococcus neoformans cells (HKC), develop two different profiles of activated T cells . CneF-CFA induces CD4+ T cells responsible for delayed-type hypersensitivity (DTH) reactivity and for amplification of the anticryptococcal DTH response, whereas HKC induce CD4+ and CD8+ T cells involved in anticryptococcal DTH reactivity and activated T cells which directly kill C . neoformans cells . The main purpose of this study was to assess the level of protection afforded by each of the two different T-cell profiles against challenge with viable C . neoformans cells, thereby identifying which activated T-cell profile provides better protection . CBA/J mice immunized with CneF-CFA had significantly better protective responses, based on better clearance of C . neoformans from tissues, on longer survival times, and on fewer and smaller lesions in the brain, than HKC-immunized mice or control mice similarly infected with C . neoformans . Both immunization protocols induced an anticryptococcal DTH response, but neither induced serum antibodies to glucuronoxylmannan, so the protection observed in the CneF-CFA immunized mice was due to the activated T-cell profile induced by that protocol . HKC-immunized mice, which displayed no greater protection than controls, did not have the amplifier cells . Based on our findings, we propose that the protective anticryptococcal T cells are the CD4+ T cells which have been shown to be responsible for DTH reactivity and/or the CD4+ T cells which amplify the DTH response and which have been previously shown to produce high levels of gamma interferon and interleukin 2 . Our results imply that there are protective and nonprotective cell-mediated immune responses and highlight the complexity of the immune response to C . neoformans antigens.

J Clin Microbiol, 1998 Apr, 36(4), 949 - 54
Identification and purification of specific Penicillium marneffei antigens and their recognition by human immune sera; Jeavons L et al.; Disseminated infection with the dimorphic pathogenic fungus Penicillium marneffei is increasingly seen among patients with AIDS in southeast Asian countries . Previous studies have demonstrated the presence of humoral immune responses to this fungus in patient sera; we have confirmed this work using sera from P . marneffei-infected patients (n = 21) to develop Western blots of P . marneffei cytoplasmic yeast antigen (CYA) . P . marneffei CYA was then partially purified by liquid isoelectric focusing, and fractions were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting . Immunoenzyme development of the Western blots with pooled sera from patients with P . marneffei infection and with pooled sera from patients with aspergillosis (n = 20), candidiasis (n = 10), cryptococcosis (n = 9), and histoplasmosis (n = 11) revealed three antigens with relative molecular masses of 61, 54, and 50 kDa . These antigens were specifically recognized by the pooled sera from the P . marneffei-infected patients . The 61- and 54-kDa antigens were subsequently purified to homogeneity by preparative gel electrophoresis, and the 50-kDa antigen was partially purified by the same technique . N-terminal amino acid sequencing revealed that the 61-kDa antigen had a strong homology (87% identity) with the antioxidant enzyme catalase . The three antigens were then subjected to SDS-PAGE and Western blotting and to immunoenzyme development with individual patient sera; sera from 86% of P . marneffei-infected patients recognized the 61-kDa antigen, sera from 71% recognized the 54-kDa antigen, and sera from 48% recognized the 50-kDa antigen . These specifically recognized antigens are the first to be purified from P . marneffei and can be used either singly or in combination to detect antibody responses in a large percentage of individuals infected with P . marneffei.

J Clin Microbiol, 1998 Apr, 36(4), 926 - 30
Comparative evaluation of FUNGITEST and broth microdilution methods for antifungal drug susceptibility testing of Candida species and Cryptococcus neoformans; Davey KG et al.; The FUNGITEST method (Sanofi Diagnostics Pasteur, Paris, France) is a microplate-based procedure for the breakpoint testing of six antifungal agents (amphotericin B, flucytosine, fluconazole, itraconazole, ketoconazole, and miconazole) . We compared the FUNGITEST method with a broth microdilution test, performed according to National Committee for Clinical Laboratory Standards document M27-A guidelines, for determining the in vitro susceptibilities of 180 isolates of Candida spp . (50 C . albicans, 50 C . glabrata, 10 C . kefyr, 20 C . krusei, 10 C . lusitaniae, 20 C . parapsilosis, and 20 C . tropicalis isolates) and 20 isolates of Cryptococcus neoformans . Overall, there was 100% agreement between the methods for amphotericin B, 95% agreement for flucytosine, 84% agreement for miconazole, 83% agreement for itraconazole, 77% agreement for ketoconazole, and 76% agreement for fluconazole . The overall agreement between the methods exceeded 80% for all species tested with the exception of C . glabrata (71% agreement) . The poorest agreement between the results for individual agents was seen with C . glabrata (38% for fluconazole, 44% for ketoconazole, and 56% for itraconazole) and C . tropicalis (50% for miconazole) . The FUNGITEST method misclassified as susceptible 2 of 12 (16.6%) fluconazole-resistant isolates, 2 of 10 (20%) itraconazole-resistant isolates, and 4 of 8 (50%) ketoconazole-resistant isolates of several Candida spp . Further development of the FUNGITEST procedure will be required before it can be recommended as an alternative method for the susceptibility testing of Candida spp . or C . neoformans.

What Is Protein?, What Is Biofilm?, What Is Growth Medium?, What Is Molecular Biology?, What Is Antibiotic?, r, Microbe, o, Bacteria, i, Microorganism, n, Microbiology, c, Microbes, i, Escherichia coli, e, Escherichia coli, i, Escherichia coli, o, Penicillin, e, Microorganisms, i, S. cerevisiae, o, Corynebacterium, e, S. cerevisiae, a, Culture medium, o, Micrococci, i, S. cerevisiae, e, Escherichia coli, c, Haemophilus, c, Typhus, i, Rhodococci, r, Erwinia, a, Corynebacter, o, Salmonella typhimurium, n, Botulism, s, Bactericidal, n, Fermentations

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005